U.S. patent application number 17/047961 was filed with the patent office on 2021-06-03 for quinoline derivatives as inhibitors of axl/mer rtk and csf1r.
The applicant listed for this patent is QURIENT CO., LTD.. Invention is credited to YEEJIN JEON, HWAN KYU KANG, JAESEUNG KIM, KIYEAN NAM, DONGSIK PARK, YEONG-IN YANG.
Application Number | 20210163448 17/047961 |
Document ID | / |
Family ID | 1000005403078 |
Filed Date | 2021-06-03 |
United States Patent
Application |
20210163448 |
Kind Code |
A1 |
NAM; KIYEAN ; et
al. |
June 3, 2021 |
QUINOLINE DERIVATIVES AS INHIBITORS OF AXL/MER RTK AND CSF1R
Abstract
The present invention relates to quinoline derivatives which are
inhibitors for Axl/Mer RTK (receptor tyrosine kinase) and CSF1R
(colony stimulating factor 1 receptor). These compounds are
suitable for the treatment of disorders associated with,
accompanied by, caused by or induced by Axl/Mer RTK and CSF1R, in
particular a hyperfunction thereof. The compounds are suitable for
the treatment of hyperproliferative disorders, such as cancer,
particularly immune-suppressive cancer (such as those cancers with
an immunosuppression of innate immunity in a tumor microenvironment
(TME), refractory cancer and cancer metastases. They are also
useful in the treatment of inflammatory diseases and/or
neurodegenerative diseases.
Inventors: |
NAM; KIYEAN; (GYEONGGI-DO,
KR) ; KIM; JAESEUNG; (SEOUL, KR) ; PARK;
DONGSIK; (GYEONGGI-DO, KR) ; JEON; YEEJIN;
(GYEONGGI-DO, KR) ; YANG; YEONG-IN; (GYEONGGI-DO,
KR) ; KANG; HWAN KYU; (GYEONGGI-DO, KR) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
QURIENT CO., LTD. |
GYEONGGI-DO |
|
KR |
|
|
Family ID: |
1000005403078 |
Appl. No.: |
17/047961 |
Filed: |
May 31, 2019 |
PCT Filed: |
May 31, 2019 |
PCT NO: |
PCT/EP2019/064214 |
371 Date: |
October 15, 2020 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
|
|
62677902 |
May 30, 2018 |
|
|
|
Current U.S.
Class: |
1/1 |
Current CPC
Class: |
C07D 405/14 20130101;
C07D 401/12 20130101; C07D 409/14 20130101; C07D 401/14
20130101 |
International
Class: |
C07D 401/14 20060101
C07D401/14; C07D 405/14 20060101 C07D405/14; C07D 401/12 20060101
C07D401/12; C07D 409/14 20060101 C07D409/14 |
Claims
1. A compound having the general formula I: ##STR00388## wherein
X.sup.1 is, independently at each occurrence, selected from
CR.sup.3 and N; X.sup.2 is, independently at each occurrence,
selected from CR.sup.4 and N; n is, independently at each
occurrence, selected from 0, 1 and 2; A is, at each occurrence,
independently selected from any structure as depicted in the
following group W; ##STR00389## R.sup.1 is, at each occurrence,
independently selected from the group consisting of hydrogen; C1-C6
alkyl; C1-C6 alkyl substituted with one or two of OR.sup.5 and
NR.sup.5R.sup.6; C3-C10 cycloalkyl; C1-C4 haloalkyl; and
--(C.dbd.O)R.sup.5; any of which is optionally substituted; R.sup.2
is, at each occurrence, independently selected from the group
consisting of C1-C6 alkyl; C3-C10 cycloalkyl; C1-C4 haloalkyl;
--NR.sup.7R.sup.8; and --OR.sup.8; any of which is optionally
substituted; R.sup.3 and R.sup.4 are, at each occurrence,
independently selected from the group consisting of hydrogen;
halogen; C1-C3 alkyl; OR.sup.5; and C1-C4 haloalkyl; any of which
is optionally substituted; R.sup.5 and R.sup.6 are, at each
occurrence, independently selected from the group consisting of
hydrogen; C1-C6 alkyl; C3-C10 cycloalkyl; and C1-C4 haloalkyl; any
of which is optionally substituted; R.sup.7 is, at each occurrence,
independently selected from the group consisting of hydrogen; C1-C6
alkyl; C1-C6 alkyl substituted with one or two of OR.sup.5 and
NR.sup.5R.sup.6; C3-C10 cycloalkyl; and C1-C4 haloalkyl; any of
which is optionally substituted; R.sup.8 is, at each occurrence,
independently selected from the group consisting of hydrogen;
--CH(CH.sub.3).sub.2; --C(CH.sub.3).sub.3; C3-C10 cycloalkyl;
C3-C10 heterocycloalkyl; C1-C4 haloalkyl; C1-C6 alkyl substituted
with one or two of OR.sup.5 and NR.sup.5R.sup.6; and C1-C6 alkyl
substituted with one or two of C3-C10 cycloalkyl, C3-C10
heterocycloalkyl and C1-C4 haloalkyl; any of which is optionally
substituted; Z.sup.1 is, at each occurrence, independently selected
from the group consisting of hydrogen; C1-C6 alkyl; C1-C6 alkyl
substituted with one or two of (.dbd.O), CN, OR.sup.5 and
NR.sup.5R.sup.6; C3-C10 cycloalkyl; C3-C10 cycloalkyl substituted
with one or several of halogen, OR.sup.7 and NR.sup.9R.sup.10;
C3-C10 heterocycloalkyl; C3-C10 heterocycloalkyl substituted with
one or several of halogen, C1-C6 alkyl, C3-C10 cycloalkyl and C1-C4
haloalkyl; and C1-C4 haloalkyl; R.sup.9 and R.sup.10 are, at each
occurrence, independently selected from the group consisting of
hydrogen; C1-C6 alkyl; C3-C10 cycloalkyl; and C1-C4 haloalkyl; any
of which is optionally substituted; R.sup.11 and R.sup.12 are, at
each occurrence, independently selected from the group consisting
of C1-C6 alkyl; C3-C10 cycloalkyl; C3-C10 heterocycloalkyl; and
C1-C4 haloalkyl; any of which is optionally substituted; and
pharmaceutically acceptable salts thereof.
2. The compound according to claim 1, having the general formula
II: ##STR00390## wherein R.sup.1, R.sup.2, R.sup.3, R.sup.11,
R.sup.12, Z.sup.1, X.sup.1, X.sup.2 and n are as defined in claim
1; and pharmaceutically acceptable salts thereof.
3. The compound according to claim 1, having the general formula
III: ##STR00391## wherein R.sup.1, R.sup.2, R.sup.3, R.sup.4,
Z.sup.1, X.sup.1, X.sup.2 and n are as defined in claim 1; and
pharmaceutically acceptable salts thereof, wherein, R.sup.3 and
R.sup.4 are, at each occurrence, independently selected from the
group consisting of hydrogen; halogen; and C1-C3 alkyl, which is
optionally substituted; R.sup.8 is, at each occurrence,
independently selected from the group consisting of hydrogen;
--CH(CH.sub.3).sub.2; --C(CH.sub.3).sub.3; C3-C10 cycloalkyl; C1-C4
haloalkyl; C1-C6 alkyl substituted with one or two of OR.sup.5 and
NR.sup.5R.sup.6; and C1-C6 alkyl substituted with one or two of
C3-C10 cycloalkyl and C1-C4 haloalkyl; any of which is optionally
substituted; and Z.sup.1 is, at each occurrence, independently
selected from the group consisting of hydrogen; C1-C6 alkyl; C1-C6
alkyl substituted with one or two of OR.sup.5 and NR.sup.5R.sup.6;
C3-C10 cycloalkyl; C3-C10 cycloalkyl substituted with one or
several of halogen, OR.sup.7 and NR.sup.9R.sup.10; C3-C10
heterocycloalkyl; C3-C10 heterocycloalkyl substituted with one or
several of halogen, C1-C6 alkyl, C3-C10 cycloalkyl and C1-C4
haloalkyl; and C1-C4 haloalkyl; and pharmaceutically acceptable
salts thereof.
4. The compound according to claim 1, having the general formula
IV: ##STR00392## wherein R.sup.1, R.sup.2, R.sup.3, R.sup.4,
Z.sup.1, X.sup.2 and n are as defined in claim 1; and
pharmaceutically acceptable salts thereof.
5. The compound according to claim 1, having the general formula V:
##STR00393## wherein R.sup.1, R.sup.2, R.sup.3, R.sup.4, Z.sup.1
and n are as defined in claim 1; wherein, R.sup.3 and R.sup.4 are
hydrogen; and pharmaceutically acceptable salts thereof.
6. The compound according to claim 1, wherein n=0 or 1, and Z.sup.1
is selected from methyl; ethyl; propyl; isopropyl; C3 cycloalkyl;
C3-C10 heterocycloalkyl; and C1-C6 alkyl substituted with one or
two of OR.sup.5 and NR.sup.5R.sup.6; and pharmaceutically
acceptable salts thereof.
7. The compound according to claim 1, wherein R.sup.2 is OR.sup.8,
and R.sup.8 is selected from --CH(CH.sub.3).sub.2;
--C(CH.sub.3).sub.3; C1-C4 haloalkyl; C1-C6 alkyl substituted with
one or two of C3-C10 cycloalkyl, C3-C10 heterocycloalkyl and C1-C4
haloalkyl; and C1-C6 alkyl substituted with one of trifluoromethyl,
difluoromethyl, fluoromethyl, trifluoroethyl, difluoroethyl,
fluoroethyl, trifluoropropyl, difluoropropyl, fluoropropyl,
trifluoroisopropyl, difluoroisopropyl, and fluoroisopropyl; and
pharmaceutically acceptable salts thereof.
8. The compound according to claim 1, wherein n=0 or 1, and Z.sup.1
is selected from methyl; ethyl; propyl; isopropyl; C3 cycloalkyl;
C3-C10 heterocycloalkyl; C1-C6 alkyl substituted with one or two of
OR.sup.5 and NR.sup.5R.sup.6; wherein R.sup.5 and R.sup.6 are, at
each occurrence, independently selected from the group consisting
of hydrogen; C1-C6 alkyl; C3-C10 cycloalkyl; and C1-C4 haloalkyl;
any of which is optionally substituted; and R.sup.2 is OR.sup.8,
and R.sup.8 is selected from --CH(CH.sub.3).sub.2;
--C(CH.sub.3).sub.3; C1-C4 haloalkyl; C1-C6 alkyl substituted with
one or two of C3-C10 cycloalkyl, C3-C10 heterocycloalkyl and C1-C4
haloalkyl; and C1-C6 alkyl substituted with one of trifluoromethyl,
difluoromethyl, fluoromethyl, trifluoroethyl, difluoroethyl,
fluoroethyl, trifluoropropyl, difluoropropyl, fluoropropyl,
trifluoroisopropyl, difluoroisopropyl, and fluoroisopropyl, and
pharmaceutically acceptable salts thereof.
9. The compound according to claim 1, wherein R.sup.2 is OR.sup.8,
and R.sup.8 is selected from one of trifluoromethyl,
difluoromethyl, fluoromethyl, trifluoroethyl, difluoroethyl,
fluoroethyl, trifluoropropyl, difluoropropyl, fluoropropyl,
trifluoroisopropyl, difluoroisopropyl, and fluoroisopropyl; or
C1-C6 alkyl substituted with one of trifluoromethyl,
difluoromethyl, fluoromethyl, trifluoroethyl, difluoroethyl,
fluoroethyl, trifluoropropyl, difluoropropyl, fluoropropyl,
trifluoroisopropyl, difluoroisopropyl, and fluoroisopropyl
10. The compound according to claim 1, wherein n=0 or 1; and
Z.sup.1 is selected from methyl; ethyl; propyl; isopropyl; C3
cycloalkyl; C4 cycloalkyl; and C5 cycloalkyl.
11. The compound according to claim 1, wherein n=0 or 1; Z.sup.1 is
selected from methyl; ethyl; propyl; isopropyl; C3 cycloalkyl; C4
cycloalkyl; and C5 cycloalkyl; and R.sup.2 is OR.sup.8; and R.sup.8
is selected from one of trifluoromethyl, difluoromethyl,
fluoromethyl, trifluoroethyl, difluoroethyl, fluoroethyl,
trifluoropropyl, difluoropropyl, fluoropropyl, trifluoroisopropyl,
difluoroisopropyl, and fluoroisopropyl; or C1-C6 alkyl substituted
with one of trifluoromethyl, difluoromethyl, fluoromethyl,
trifluoroethyl, difluoroethyl, fluoroethyl, trifluoropropyl,
difluoropropyl, fluoropropyl, trifluoroisopropyl,
difluoroisopropyl, and fluoroisopropyl
12. The compound according to claim 1 having one of the structures
as shown hereafter: TABLE-US-00010 # cpd Structure 1 ##STR00394## 2
##STR00395## 3 ##STR00396## 4 ##STR00397## 5 ##STR00398## 6
##STR00399## 7 ##STR00400## 8 ##STR00401## 9 ##STR00402## 10
##STR00403## 11 ##STR00404## 12 ##STR00405## 13 ##STR00406## 14
##STR00407## 15 ##STR00408## 16 ##STR00409## 17 ##STR00410## 18
##STR00411## 19 ##STR00412## 20 ##STR00413## 21 ##STR00414## 22
##STR00415## 23 ##STR00416## 24 ##STR00417## 25 ##STR00418## 26
##STR00419## 27 ##STR00420## 28 ##STR00421## 29 ##STR00422## 30
##STR00423## 31 ##STR00424## 32 ##STR00425## 33 ##STR00426## 34
##STR00427## 35 ##STR00428## 36 ##STR00429## 37 ##STR00430## 38
##STR00431## 39 ##STR00432## 40 ##STR00433## 41 ##STR00434## 42
##STR00435## 43 ##STR00436## 44 ##STR00437## 45 ##STR00438## 46
##STR00439## 47 ##STR00440## 48 ##STR00441## 49 ##STR00442## 50
##STR00443## 51 ##STR00444## 52 ##STR00445## 53 ##STR00446## 54
##STR00447## 55 ##STR00448## 56 ##STR00449## 57 ##STR00450## 58
##STR00451## 59 ##STR00452## 60 ##STR00453## 61 ##STR00454## 62
##STR00455## 63 ##STR00456## 64 ##STR00457## 65 ##STR00458## 66
##STR00459## 67 ##STR00460## 68 ##STR00461## 69 ##STR00462## 70
##STR00463## 71 ##STR00464## 72 ##STR00465## 73 ##STR00466## 74
##STR00467## 75 ##STR00468## 76 ##STR00469## 77 ##STR00470## 78
##STR00471## 79 ##STR00472## 80 ##STR00473## 81 ##STR00474## 82
##STR00475## 83 ##STR00476## 84 ##STR00477## 85 ##STR00478## 86
##STR00479## 87 ##STR00480## 88 ##STR00481## 89 ##STR00482## 90
##STR00483## 91 ##STR00484## 92 ##STR00485## 93 ##STR00486## 94
##STR00487## 95 ##STR00488## 96 ##STR00489## 97 ##STR00490## 98
##STR00491## 99 ##STR00492## 100 ##STR00493## 101 ##STR00494## 102
##STR00495## 103 ##STR00496## 104 ##STR00497## 105 ##STR00498## 106
##STR00499## 107 ##STR00500## 108 ##STR00501## 109 ##STR00502## 110
##STR00503## 111 ##STR00504## 112 ##STR00505## 113 ##STR00506## 114
##STR00507## 115 ##STR00508## 116 ##STR00509## 117 ##STR00510## 118
##STR00511## 119 ##STR00512## 120 ##STR00513## 121 ##STR00514## 122
##STR00515## 123 ##STR00516##
124 ##STR00517## 125 ##STR00518## 126 ##STR00519## 127 ##STR00520##
128 ##STR00521## 129 ##STR00522## 130 ##STR00523## 131 ##STR00524##
132 ##STR00525## 133 ##STR00526## 134 ##STR00527## 135 ##STR00528##
136 ##STR00529## 137 ##STR00530## 138 ##STR00531## 139 ##STR00532##
140 ##STR00533## 141 ##STR00534## 142 ##STR00535## 143 ##STR00536##
144 ##STR00537## 145 ##STR00538## 146 ##STR00539## 147 ##STR00540##
148 ##STR00541## 149 ##STR00542## 150 ##STR00543## 151 ##STR00544##
152 ##STR00545## 153 ##STR00546## 154 ##STR00547## 155 ##STR00548##
156 ##STR00549## 157 ##STR00550## 158 ##STR00551## 159 ##STR00552##
160 ##STR00553## 161 ##STR00554## 162 ##STR00555## 163 ##STR00556##
164 ##STR00557## 165 ##STR00558## 166 ##STR00559## 167 ##STR00560##
168 ##STR00561## 169 ##STR00562## 170 ##STR00563## 171 ##STR00564##
172 ##STR00565## 173 ##STR00566## 174 ##STR00567## 175 ##STR00568##
176 ##STR00569## 177 ##STR00570## 178 ##STR00571## 179
##STR00572##
13. A composition comprising at least one compound according to
claim 1, together with at least one pharmaceutically acceptable
carrier, excipient and/or diluent.
14. The composition according to claim 13, further comprising at
least one other pharmaceutically active agent.
15. (canceled)
16. A method for the treatment of a disorder associated with,
accompanied by, caused by or induced by a Axl/Mer and CSF1R
receptor tyrosine kinase wherein said method comprises
administering, to subject in need of such treatment, a compound of
claim 1.
17. The method according to claim 16, wherein said disorder is
selected from hyperproliferative disorders, inflammatory disorders
and neurodegenerative disorders.
18. The method according to claim 17, wherein said
hyperproliferative disorder is a cancer selected from
adenocarcinoma, acoustic neuroma, acute lymphoblastic leukemia,
acute myeloid leukemia, adrenocortical carcinoma, aids-related
cancers, aids-related lymphoma, anal cancer, appendix cancer,
astrocytomas, atypical teratoid/rhabdoid tumor, ampullary
carcinoma, basal cell carcinoma, bile duct cancer, bladder cancer,
bone cancer, osteosarcoma and malignant fibrous histiocytoma, brain
stem glioma, brain tumor, central nervous system atypical
teratoid/rhabdoid tumor, craniopharyngioma, ependymoblastoma,
ependymoma, medulloblastoma, medulloepithelioma, pineal parenchymal
tumors of intermediate differentiation, supratentorial primitive
neuroectodermal tumors and pineoblastoma, brain and spinal cord
tumors, breast cancer, urachal tumors, burkitt lymphoma, carcinoid
tumor, choroidal melanoma, gastrointestinal cancer, central nervous
system lymphoma, cervical cancer, corpus cancer, chordoma, chronic
lymphocytic leukemia, chronic myelogenous leukemia, chronic
myeloproliferative disorders, colon cancer, colorectal cancer,
cutaneous t-cell lymphoma, desmoid tumor, mycosis fungoides,
endometrial cancer, esophageal cancer, esthesioneuroblastoma, Ewing
sarcoma family of tumors, extracranial germ cell tumor,
extragonadal germ cell tumor, extrahepatic bile duct cancer, ear
tumors, intraocular melanoma, retinoblastoma, gallbladder cancer,
gastric cancer, gastrointestinal carcinoid tumor, gastrointestinal
stromal tumor, gastrointestinal stromal cell tumor, gynecologic
tumors, ovarian germ cell tumor, gestational trophoblastic tumor,
glioma, gallbladder carcinomas, hairy cell leukemia, head and neck
cancer, heart cancer, hepatocellular cancer, histiocytosis,
hypopharyngeal cancer, hematologic neoplasias, islet cell tumors
(endocrine pancreas), renal cell cancer, kidney cancer, Langerhans
cell histiocytosis, laryngeal cancer, leukemia, lip and oral cavity
cancer, liver cancer, lung cancer, non-small cell lung cancer,
small intestinal tumors, small cell lung cancer, hodgkin lymphoma,
non-hodgkin lymphoma, primary central nervous system lymphoma,
macroglobulinemia, malignant fibrous histiocytoma of bone and
osteosarcoma, melanoma, merkel cell carcinoma, mesothelioma,
metastatic squamous neck cancer with occult primary, spinalioms,
multiple endocrine neoplasia syndromes, myelodysplastic syndromes,
myelodysplastic/myeloproliferative neoplasms, myeloid leukemia,
multiple myeloma, myeloproliferative disorders, nasal cavity and
paranasal sinus cancer, nasopharyngeal cancer, neuroblastoma, oral
cavity cancer, oropharyngeal cancer, osteosarcoma and malignant
fibrous histiocytoma of bone, ovarian cancer, ovarian epithelial
cancer, ovarian low malignant potential tumor, oligodendroglioma,
plasmacytomas, pancreatic cancer, papillomatosis, parathyroid
cancer, penile cancer, pharyngeal cancer, pituitary tumor, plasma
cell neoplasm/multiple myeloma, pleuropulmonary blastoma, prostate
cancer, rectal cancer, renal cell cancer, transitional cell cancer,
respiratory tract cancer, rhabdomyosarcoma, salivary gland cancer,
sarcoma, skin testis cancer, ewing sarcoma, kaposi sarcoma, uterine
sarcoma, non-melanoma skin cancer, melanoma skin cancer, skin
carcinoma, small intestine cancer, soft tissue sarcoma, squamous
cell carcinoma, squamous neck cancer, stomach cancer, soft tissue
tumors, testicular cancer, throat cancer, thymoma and thymic
carcinoma, thyroid cancer, transitional cell cancer of the renal
pelvis and ureter, trophoblastic tumor, testicle cancer,
gestational cancer, urologic tumors, ureter and renal pelvis
cancer, urethral cancer, urothelial carcinoma, uterine cancer,
vaginal cancer, vulvar cancer, waldenstrom macroglobulinemia and
wilms tumor, tumors that cause effusions in potential spaces of the
body, pleural effusions, pericardial effusions, peritoneal effusion
aka ascites, giant cell tumor (GCT), GCT of bone, pigmented
villonodular synovitis (PVNS), and tenosynovial giant cell tumor
(TGCT), TGCT of tendon sheath (TGCT-TS).
19. The method according to claim 17, wherein said inflammatory
disorder is selected from osteoarthritis, inflammatory bowel
syndrome, transplant rejection, systemic lupus erythematosis,
ulcerative colitis, crohn's disease, chronic obstructive pulmonary
disease, emphysema, Kawasaki's Disease, hemophagocytic syndrome
(macrophage activation syndrome), multicentric
reticulohistiocytosis, atherosclerosis, primary progressive
multiple sclerosis, tenpsy Type I diabetes, Type II diabetes,
insulin resistance, hyperglycemia, obesity, lipolysis,
hypereosinophilia, osteoporosis, increased risk of fracture,
Paget's disease, hypercalcemia, infection mediated osteolysis (e.g.
osteomyelitis), peri-prosthetic or wear-debris-mediated osteolysis,
endometriosis, inflammatory pain, chronic pain, and bone pain.
20. The method according to claim 17, wherein said
neurodegenerative disorder is selected from Binswanger type
dementia, prosencephaly, microcephaly, cerebral palsy, congenital
hydrocephalus, abdominal dropsy, progress supranuclear palsy,
glaucoma, Wilson disease, Alzheimer's disease and other dementias,
Parkinson's disease (PD) and PD-related disorders, multi infarct
dementia, Frontotemporal dementia, pseudo-dementia, Prion disease,
Motor neuron diseases, Huntington's disease, spinocerebellar
ataxia, and spinal muscular atrophy.
21. The method according to claim 16, which further comprises the
administration of radiation therapy, chemotherapy agents, targeted
drugs and/or immune check point inhibitor drugs.
22. (canceled)
Description
[0001] The present invention relates to quinoline derivatives which
are inhibitors for Axl/Mer RTK (receptor tyrosine kinase) and CSF1R
(colony stimulating factor 1 receptor). These compounds are
suitable for the treatment of disorders associated with,
accompanied by, caused by or induced by Axl/Mer RTK and CSF1R, in
particular a hyperfunction thereof. The compounds are suitable for
the treatment of hyperproliferative disorders, such as cancer,
particularly immune-suppressive cancer (such as those cancers with
an immunosuppression of innate immunity in a tumor microenvironment
(TME), refractory cancer and cancer metastases. They are also
useful in the treatment of inflammatory diseases and/or
neurodegenerative diseases.
BACKGROUND OF THE INVENTION
[0002] Axl/Mer receptor tyrosine kinase (Axl/Mer RTK) are member of
the TAM (Tyrosine, Axl, Mer) receptor tyrosine kinases. They are
characterised by extracellular domain, consisting two
immunoglobulin-like domains followed by two fibronectin type-3-like
domain. The activation of the Axl/Mer occurs by its cognate protein
ligand, growth arrest specific 6 (Gas6) and protein S (Pros1),
respectively.
[0003] Axl/Mer RTK is already known to regulate cell growth,
differentiation and survival, but in recent years Axl/Mer RTK has
become a promising target for cancer immunotherapy and is known to
be a regulator of immune homeostasis. Guided by their cognate
ligands Gas6 and Pros1, these receptors ensure the resolution of
inflammation by dampening the activation of innate cells as well as
by restoring tissue function through promotion of tissue repair and
clearance of apoptotic cells in normal condition (Paolino M et al.,
Cancers (Basel). 2016 Oct. 21; 8(10) pii: E97).
[0004] However, in the tumor microenvironment (TME), activation of
Axl/Mer receptors can lead to immune evasion via Axl/Mer RTK driven
phagocytosis of apoptotic cells, negative regulation of the immune
response and subsequent inhibition of T-cell priming (Zagorska A et
al., Nat Immunol. 2014 October; 15(10):920-8). In addition, Axl/Mer
RTK play a role in modulating the cytokine milieu of the tumor to
limit effective CD8+ T cell recruitment and to polarize macrophages
toward and M2 anti-inflammatory state which plays important role in
enhancing tumor progression. (Akalu Y T et al., Immunol Rev. 2017
March; 276(1):165-177). Moreover, inhibition of TLR pathway by
Axl/Mer RTK in DCs triggers suppression of CD8+ cytotoxic T cell
population, which can increase anti-tumor immunity. In addition
Activation of TAM family is reported to downregulate TLR
signaling-mediated inflammatory response through increase in SOCS1
and 3 expressions.
[0005] Axl/Mer receptors are also reported to negatively regulate
activation of NK cells through regulating Cbl/b as well as
suppresses the IFN gamma cytokine production of activated NK cells.
Accordingly, inhibiting Axl/Mer receptors are possible with new
immunotherapy approaches for various cancers patients by priming
host immune response against tumour cell invasion (Davra V et al.,
Cancers (Basel). 2016 Nov. 29; 8(12). pii: E107).
[0006] Colony stimulating factor 1 receptor (CSF1R) is also cell
surface homodimeric type III receptor tyrosine kinase by macrophage
and other cells of the myeloid lineage that is encoded by the c-fms
proto-oncogene. The activation of the CSF1R occurs by bound by its
ligand. CSF1 and IL-34 (Hume D A et al., Blood. 2012 Feb. 23;
119(8):1810-20).
[0007] CSF1R is known to regulate the differentiation of myeloid
progenitors into heterogeneous populations of monocytes,
macrophages, dendritic cells (DC) and bone-resorbing osteoclasts.
In addition, activated CSF1R promotes the survival, proliferation,
differentiation and chemotaxis of differentiated macrophages
(Geissmann F et al., Science. 2010 Feb. 5; 327(5966):656-61).
[0008] Based on role of CSF1R in immune cells, various approaches
targeting either the CSF1R or its ligands is developing against
immunotherapy and cancers and currently it in clinical stage.
SUMMARY OF THE INVENTION
[0009] It is an object of the present invention to provide
compounds and/or pharmaceutically acceptable salts thereof which
can be used as pharmaceutically active agents, especially for
treatment of cell proliferative diseases like cancer, but also
inflammatory diseases and/or neurodegenerative diseases, as well as
compositions comprising at least one of those compounds and/or
pharmaceutically acceptable salts thereof as pharmaceutically
active ingredients.
[0010] In a first aspect, the present invention relates to a
compound having the general formula I:
##STR00001## [0011] wherein [0012] X.sup.1 is, independently at
each occurrence, selected from CR.sup.3 and N; [0013] X.sup.2 is,
independently at each occurrence, selected from CR.sup.4 and N;
[0014] n is, independently at each occurrence, selected from 0, 1
and 2; [0015] A is, at each occurrence, independently selected from
any structure as depicted in the following group W;
[0015] ##STR00002## ##STR00003## [0016] R.sup.1 is, at each
occurrence, independently selected from the group consisting of
hydrogen; C1-C6 alkyl; C1-C6 alkyl substituted with one or two of
OR.sup.5 and NR.sup.5R.sup.6; C3-C10 cycloalkyl; C1-C4 haloalkyl;
--(C.dbd.O)R.sup.5; any of which is optionally substituted; [0017]
R.sup.2 is, at each occurrence, independently selected from the
group consisting of C1-C6 alkyl; C3-C10 cycloalkyl; C1-C4
haloalkyl; --NR.sup.7R.sup.8; --OR.sup.8; any of which is
optionally substituted; [0018] R.sup.3 and R.sup.4 are, at each
occurrence, independently selected from the group consisting of
hydrogen; halogen, e.g Cl or F; C1-C3 alkyl; OR.sup.5; C1-C4
haloalkyl; any of which is optionally substituted; [0019] R.sup.5
and R.sup.6 are, at each occurrence, independently selected from
the group consisting of hydrogen; C1-C6 alkyl; C3-C10 cycloalkyl;
C1-C4 haloalkyl; any of which is optionally substituted; [0020]
R.sup.7 is, at each occurrence, independently selected from the
group consisting of hydrogen; C1-C6 alkyl; C1-C6 alkyl substituted
with one or two of OR.sup.5 and NR.sup.5R.sup.6; C3-C10 cycloalkyl;
C1-C4 haloalkyl; any of which is optionally substituted; [0021]
R.sup.8 is, at each occurrence, independently selected from the
group consisting of hydrogen; --CH(CH.sub.3).sub.2;
--C(CH.sub.3).sub.3; C3-C10 cycloalkyl; C3-C10 heterocycloalkyl;
C1-C4 haloalkyl; C1-C6 alkyl substituted with one or two of
OR.sup.5 and NR.sup.5R.sup.6; and C1-C6 alkyl substituted with one
or two of C3-C10 cycloalkyl, C3-C10 heterocycloalkyl and C1-C4
haloalkyl; any of which is optionally substituted; [0022] Z.sup.1
is, at each occurrence, independently selected from the group
consisting of hydrogen; C1-C6 alkyl; C1-C6 alkyl substituted with
one or two of (.dbd.O), CN, OR.sup.5 and NR.sup.5R.sup.6; C3-C10
cycloalkyl; C3-C10 cycloalkyl substituted with one or several of
halogen, OR.sup.7 and NR.sup.9R.sup.10; C3-C10 heterocycloalkyl;
C3-C10 heterocycloalkyl substituted with one or several of halogen,
C1-C6 alkyl, C3-C10 cycloalkyl and C1-C4 haloalkyl; C1-C4
haloalkyl; [0023] R.sup.9 and R.sup.10 are, at each occurrence,
independently selected from the group consisting of hydrogen; C1-C6
alkyl; C3-C10 cycloalkyl; C1-C4 haloalkyl; any of which is
optionally substituted; [0024] R.sup.11 and R.sup.12 are, at each
occurrence, independently selected from the group consisting of
C1-C6 alkyl; C3-C10 cycloalkyl; C3-C10 heterocycloalkyl; C1-C4
haloalkyl; any of which is optionally substituted; [0025] and
pharmaceutically acceptable salts thereof.
[0026] In one embodiment, the compound according to the present
invention has the general formula II:
##STR00004## [0027] wherein [0028] R.sup.1, R.sup.2, R.sup.3,
R.sup.4, R.sup.11, R.sup.12, Z.sup.1, X.sup.1, X.sup.2 and n are as
defined above; and [0029] pharmaceutically acceptable salts
thereof.
[0030] In one embodiment, the compound according to the present
invention has the general formula III:
##STR00005## [0031] wherein [0032] R.sup.1, R.sup.2, R.sup.3,
R.sup.4, Z.sup.1, X.sup.1, X.sup.2 and n are as defined above; and
pharmaceutically acceptable salts thereof, [0033] wherein,
preferably, [0034] R.sup.3 and R.sup.4 are, at each occurrence,
independently selected from the group consisting of hydrogen;
halogen, e.g Cl or F; C1-C3 alkyl, which is optionally substituted;
[0035] R.sup.8 is, at each occurrence, independently selected from
the group consisting of hydrogen; --CH(CH.sub.3).sub.2;
--C(CH.sub.3).sub.3; C3-C10 cycloalkyl; C1-C4 haloalkyl; C1-C6
alkyl substituted with one or two of OR.sup.5 and NR.sup.5R.sup.6;
or C1-C6 alkyl substituted with one or two of C3-C10 cycloalkyl and
C1-C4 haloalkyl; any of which is optionally substituted; and [0036]
Z.sup.1 is, at each occurrence, independently selected from the
group consisting of hydrogen; C1-C6 alkyl; C1-C6 alkyl substituted
with one or two of OR.sup.5 and NR.sup.5R.sup.6; C3-C10 cycloalkyl;
C3-C10 cycloalkyl substituted with one or several of halogen,
OR.sup.7 and NR.sup.9R.sup.10; C3-C10 heterocycloalkyl; C3-C10
heterocycloalkyl substituted with one or several of halogen, C1-C6
alkyl, C3-C10 cycloalkyl and C1-C4 haloalkyl; C1-C4 haloalkyl; and
pharmaceutically acceptable salts thereof.
[0037] In one embodiment, the compound according to the present
invention has the general formula IV:
##STR00006## [0038] wherein [0039] R.sup.1, R.sup.2, R.sup.3,
R.sup.4, Z.sup.1, X.sup.2 and n are as defined above; and
pharmaceutically acceptable salts thereof.
[0040] In one embodiment, the compound according to the present
invention has the general formula V:
##STR00007## [0041] wherein [0042] R.sup.1, R.sup.2, R.sup.3,
R.sup.4, Z.sup.1 and n are as defined above; and pharmaceutically
acceptable salts thereof; [0043] wherein, preferably, [0044]
R.sup.3 and R.sup.4 are hydrogen; [0045] and pharmaceutically
acceptable salts thereof.
[0046] In one embodiment n=0 or 1, and Z.sup.1 is selected from
C1-C6 alkyl, in particular methyl, ethyl, propyl or isopropyl;
C3-C10 cycloalkyl, in particular C3 cycloalkyl; C3-C10
heterocycloalkyl; C1-C6 alkyl substituted with one or two of
OR.sup.5 and NR.sup.5R.sup.6; and pharmaceutically acceptable salts
thereof.
[0047] In one embodiment R.sup.2 is OR.sup.8, and R.sup.8 is
selected from --CH(CH.sub.3).sub.2; --C(CH.sub.3).sub.3; C1-C4
haloalkyl; C1-C6 alkyl substituted with one or two of C3-C10
cycloalkyl, C3-C10 heterocycloalkyl and C1-C4 haloalkyl; or C1-C6
alkyl substituted with C1-C4 haloalkyl, in particular one of
trifluoromethyl, difluoromethyl, fluoromethyl, trifluoroethyl,
difluoroethyl, fluoroethyl, trifluoropropyl, difluoropropyl,
fluoropropyl, trifluoroisopropyl, difluoroisopropyl, and
fluoroisopropyl and pharmaceutically acceptable salts thereof.
[0048] In one embodiment n=0 or 1, and Z.sup.1 is selected from
C1-C6 alkyl, in particular methyl, ethyl, propyl or isopropyl;
C3-C10 cycloalkyl, in particular C3 cycloalkyl; C3-C10
heterocycloalkyl; C1-C6 alkyl substituted with one or two of
OR.sup.5 and NR.sup.5R.sup.6; wherein R.sup.5 and R.sup.6 are, at
each occurrence, independently selected from the group consisting
of hydrogen; C1-C6 alkyl; C3-C10 cycloalkyl; C1-C4 haloalkyl; any
of which is optionally substituted; [0049] and [0050] R.sup.2 is
OR.sup.8, and R.sup.8 is selected from --CH(CH.sub.3).sub.2;
--C(CH.sub.3).sub.3; C1-C4 haloalkyl; C1-C6 alkyl substituted with
one or two of C3-C10 cycloalkyl, C3-C10 heterocycloalkyl and C1-C4
haloalkyl; or C1-C6 alkyl substituted with C1-C4 haloalkyl, in
particular one of trifluoromethyl, difluoromethyl, fluoromethyl,
trifluoroethyl, difluoroethyl, fluoroethyl, trifluoropropyl,
difluoropropyl, fluoropropyl, trifluoroisopropyl,
difluoroisopropyl, and fluoroisopropyl [0051] and pharmaceutically
acceptable salts thereof.
[0052] In one embodiment R.sup.2 is OR.sup.8, and R.sup.8 is
selected from C1-C4 haloalkyl, in particular one of
trifluoromethyl, difluoromethyl, fluoromethyl, trifluoroethyl,
difluoroethyl, fluoroethyl, trifluoropropyl, difluoropropyl,
fluoropropyl, trifluoroisopropyl, difluoroisopropyl, and
fluoroisopropyl; or C1-C6 alkyl substituted with C1-C4 haloalkyl,
in particular one of trifluoromethyl, difluoromethyl, fluoromethyl,
trifluoroethyl, difluoroethyl, fluoroethyl, trifluoropropyl,
difluoropropyl, fluoropropyl, trifluoroisopropyl,
difluoroisopropyl, and fluoroisopropyl
[0053] In one embodiment n=0 or 1; and Z.sup.1 is selected from
methyl; ethyl; propyl; isopropyl; C3 cycloalkyl; C4 cycloalkyl; and
C5 cycloalkyl.
[0054] In one embodiment n=0 or 1; Z.sup.1 is selected from methyl;
ethyl; propyl; isopropyl; C3 cycloalkyl; C4 cycloalkyl; and C5
cycloalkyl; and
[0055] R.sup.2 is OR.sup.8; and R.sup.8 is selected from C1-C4
haloalkyl, in particular one of trifluoromethyl, difluoromethyl,
fluoromethyl, trifluoroethyl, difluoroethyl, fluoroethyl,
trifluoropropyl, difluoropropyl, fluoropropyl, trifluoroisopropyl,
difluoroisopropyl, and fluoroisopropyl; or C1-C6 alkyl substituted
with C1-C4 haloalkyl, in particular one of trifluoromethyl,
difluoromethyl, fluoromethyl, trifluoroethyl, difluoroethyl,
fluoroethyl, trifluoropropyl, difluoropropyl, fluoropropyl,
trifluoroisopropyl, difluoroisopropyl, and fluoroisopropyl
[0056] In one embodiment, the compound according to the present
invention has one of the structures as shown hereafter:
TABLE-US-00001 # cpd Structure 1 ##STR00008## 2 ##STR00009## 3
##STR00010## 4 ##STR00011## 5 ##STR00012## 6 ##STR00013## 7
##STR00014## 8 ##STR00015## 9 ##STR00016## 10 ##STR00017## 11
##STR00018## 12 ##STR00019## 13 ##STR00020## 14 ##STR00021## 15
##STR00022## 16 ##STR00023## 17 ##STR00024## 18 ##STR00025## 19
##STR00026## 20 ##STR00027## 21 ##STR00028## 22 ##STR00029## 23
##STR00030## 24 ##STR00031## 25 ##STR00032## 26 ##STR00033## 27
##STR00034## 28 ##STR00035## 29 ##STR00036## 30 ##STR00037## 31
##STR00038## 32 ##STR00039## 33 ##STR00040## 34 ##STR00041## 35
##STR00042## 36 ##STR00043## 37 ##STR00044## 38 ##STR00045## 39
##STR00046## 40 ##STR00047## 41 ##STR00048## 42 ##STR00049## 43
##STR00050## 44 ##STR00051## 45 ##STR00052## 46 ##STR00053## 47
##STR00054## 48 ##STR00055## 49 ##STR00056## 50 ##STR00057## 51
##STR00058## 52 ##STR00059## 53 ##STR00060## 54 ##STR00061## 55
##STR00062## 56 ##STR00063## 57 ##STR00064## 58 ##STR00065## 59
##STR00066## 60 ##STR00067## 61 ##STR00068## 62 ##STR00069## 63
##STR00070## 64 ##STR00071## 65 ##STR00072## 66 ##STR00073## 67
##STR00074## 68 ##STR00075## 69 ##STR00076## 70 ##STR00077## 71
##STR00078## 72 ##STR00079## 73 ##STR00080## 74 ##STR00081## 75
##STR00082## 76 ##STR00083## 77 ##STR00084## 78 ##STR00085## 79
##STR00086## 80 ##STR00087## 81 ##STR00088## 82 ##STR00089## 83
##STR00090## 84 ##STR00091## 85 ##STR00092## 86 ##STR00093## 87
##STR00094## 88 ##STR00095## 89 ##STR00096## 90 ##STR00097## 91
##STR00098## 92 ##STR00099## 93 ##STR00100## 94 ##STR00101## 95
##STR00102## 96 ##STR00103## 97 ##STR00104## 98 ##STR00105## 99
##STR00106## 100 ##STR00107## 101 ##STR00108## 102 ##STR00109## 103
##STR00110## 104 ##STR00111## 105 ##STR00112## 106 ##STR00113## 107
##STR00114## 108 ##STR00115## 109 ##STR00116## 110 ##STR00117## 111
##STR00118## 112 ##STR00119## 113 ##STR00120## 114 ##STR00121## 115
##STR00122## 116 ##STR00123## 117 ##STR00124## 118 ##STR00125## 119
##STR00126## 120 ##STR00127## 121 ##STR00128## 122 ##STR00129## 123
##STR00130##
124 ##STR00131## 125 ##STR00132## 126 ##STR00133## 127 ##STR00134##
128 ##STR00135## 129 ##STR00136## 130 ##STR00137## 131 ##STR00138##
132 ##STR00139## 133 ##STR00140## 134 ##STR00141## 135 ##STR00142##
136 ##STR00143## 137 ##STR00144## 138 ##STR00145## 139 ##STR00146##
140 ##STR00147## 141 ##STR00148## 142 ##STR00149## 143 ##STR00150##
144 ##STR00151## 145 ##STR00152## 146 ##STR00153## 147 ##STR00154##
148 ##STR00155## 149 ##STR00156## 150 ##STR00157## 151 ##STR00158##
152 ##STR00159## 153 ##STR00160## 154 ##STR00161## 155 ##STR00162##
156 ##STR00163## 157 ##STR00164## 158 ##STR00165## 159 ##STR00166##
160 ##STR00167## 161 ##STR00168## 162 ##STR00169## 163 ##STR00170##
164 ##STR00171## 165 ##STR00172## 166 ##STR00173## 167 ##STR00174##
168 ##STR00175## 169 ##STR00176## 170 ##STR00177## 171 ##STR00178##
172 ##STR00179## 173 ##STR00180## 174 ##STR00181## 175 ##STR00182##
176 ##STR00183## 177 ##STR00184## 178 ##STR00185## 179
##STR00186##
[0057] In a further aspect, the present invention relates to a
composition comprising at least one compound according the present
invention, together with at least one pharmaceutically acceptable
carrier, excipient and/or diluent.
[0058] In one embodiment, the composition according to the present
invention further comprises at least one other pharmaceutically
active agent.
[0059] In a further aspect, the present invention relates to a
compound according to the present invention or a composition
according to the present invention, for use as a pharmaceutically
active agent, preferably for use in a method of treating a
disorder.
[0060] In a further aspect, the present invention relates to a
compound according to the present invention or the composition
according to the present invention, for use in the treatment of a
disorder associated with, accompanied by, caused by or induced by a
Axl/Mer and CSF1R receptor tyrosine kinase, in particular
associated with, accompanied by, or caused by Axl/Mer and CSF1R
(colony-stimulating factor-1-receptor), preferably associated with,
accompanied by, or caused by a hyperfunction of said Axl/Mer and a
hyperfunction of said CSF1R.
[0061] In one embodiment, said disorder is selected from
hyperproliferative disorders, inflammatory disorders and
neurodegenerative disorders.
[0062] In one embodiment, said hyperproliferative disorder is a
cancer, preferably a cancer selected from adenocarcinoma, acoustic
neuroma, acute lymphoblastic leukemia, acute myeloid leukemia,
adrenocortical carcinoma, aids-related cancers, aids-related
lymphoma, anal cancer, appendix cancer, astrocytomas, atypical
teratoid/rhabdoid tumor, ampullary carcinoma, basal cell carcinoma,
bile duct cancer, bladder cancer, bone cancer, osteosarcoma and
malignant fibrous histiocytoma, brain stem glioma, brain tumor,
central nervous system atypical teratoid/rhabdoid tumor,
craniopharyngioma, ependymoblastoma, ependymoma, medulloblastoma,
medulloepithelioma, pineal parenchymal tumors of intermediate
differentiation, supratentorial primitive neuroectodermal tumors
and pineoblastoma, brain and spinal cord tumors, breast cancer,
urachal tumors, burkitt lymphoma, carcinoid tumor, choroidal
melanoma, gastrointestinal cancer, central nervous system lymphoma,
cervical cancer, corpus cancer, chordoma, chronic lymphocytic
leukemia, chronic myelogenous leukemia, chronic myeloproliferative
disorders, colon cancer, colorectal cancer, cutaneous t-cell
lymphoma, desmoid tumor, mycosis fungoides, endometrial cancer,
esophageal cancer, esthesioneuroblastoma, Ewing sarcoma family of
tumors, extracranial germ cell tumor, extragonadal germ cell tumor,
extrahepatic bile duct cancer, ear tumors, intraocular melanoma,
retinoblastoma, gallbladder cancer, gastric cancer,
gastrointestinal carcinoid tumor, gastrointestinal stromal tumor,
gastrointestinal stromal cell tumor, gynecologic tumors, ovarian
germ cell tumor, gestational trophoblastic tumor, glioma,
gallbladder carcinomas, hairy cell leukemia, head and neck cancer,
heart cancer, hepatocellular cancer, histiocytosis, hypopharyngeal
cancer, hematologic neoplasias, islet cell tumors (endocrine
pancreas), renal cell cancer, kidney cancer, langerhans cell
histiocytosis, laryngeal cancer, leukemia, lip and oral cavity
cancer, liver cancer, lung cancer, non-small cell lung cancer,
small intestinal tumors, small cell lung cancer, hodgkin lymphoma,
non-hodgkin lymphoma, primary central nervous system lymphoma,
macroglobulinemia, malignant fibrous histiocytoma of bone and
osteosarcoma, melanoma, merkel cell carcinoma, mesothelioma,
metastatic squamous neck cancer with occult primary, spinalioms,
multiple endocrine neoplasia syndromes, myelodysplastic syndromes,
myelodysplastic/myeloproliferative neoplasms, myeloid leukemia,
multiple myeloma, myeloproliferative disorders, nasal cavity and
paranasal sinus cancer, nasopharyngeal cancer, neuroblastoma, oral
cavity cancer, oropharyngeal cancer, osteosarcoma and malignant
fibrous histiocytoma of bone, ovarian cancer, ovarian epithelial
cancer, ovarian low malignant potential tumor, oligodendroglioma,
plasmacytomas, pancreatic cancer, papillomatosis, parathyroid
cancer, penile cancer, pharyngeal cancer, pituitary tumor, plasma
cell neoplasm/multiple myeloma, pleuropulmonary blastoma, prostate
cancer, rectal cancer, renal cell cancer, transitional cell cancer,
respiratory tract cancer, rhabdomyosarcoma, salivary gland cancer,
sarcoma, skin testis cancer, ewing sarcoma, kaposi sarcoma, uterine
sarcoma, non-melanoma skin cancer, melanoma skin cancer, skin
carcinoma, small intestine cancer, soft tissue sarcoma, squamous
cell carcinoma, squamous neck cancer, stomach cancer, soft tissue
tumors, testicular cancer, throat cancer, thymoma and thymic
carcinoma, thyroid cancer, transitional cell cancer of the renal
pelvis and ureter, trophoblastic tumor, testicle cancer,
gestational cancer, urologic tumors, ureter and renal pelvis
cancer, urethral cancer, urothelial carcinoma, uterine cancer,
vaginal cancer, vulvar cancer, waldenstrom macroglobulinemia and
wilms tumor, tumors that cause effusions in potential spaces of the
body, pleural effusions, pericardial effusions, peritoneal effusion
aka ascites, giant cell tumor (GCT), GCT of bone, pigmented
villonodular synovitis (PVNS), tenosynovial giant cell tumor
(TGCT), TGCT of tendon sheath (TGCT-TS).
[0063] In one embodiment, said inflammatory disorder is selected
from osteoarthritis, inflammatory bowel syndrome, transplant
rejection, systemic lupus erythematosis, ulcerative colitis,
crohn's disease, chronic obstructive pulmonary disease, emphysema,
Kawasaki's Disease, hemophagocytic syndrome (macrophage activation
syndrome), multicentric reticulohistiocytosis, atherosclerosis,
primary progressive multiple sclerosis, tenpsy Type I diabetes,
Type Il diabetes, insulin resistance, hyperglycemia, obesity,
lipolysis, hypereosinophilia, osteoporosis, increased risk of
fracture, Paget's disease, hypercalcemia, infection mediated
osteolysis (e.g. osteomyelitis), peri-prosthetic or
wear-debris-mediated osteolysis, endometriosis, inflammatory pain,
chronic pain, and bone pain.
[0064] In one embodiment, said neurodegenerative disorder is
selected from Binswanger type dementia, prosencephaly,
microcephaly, cerebral palsy, congenital hydrocephalus, abdominal
dropsy, progress supranuclear palsy, glaucoma, Wilson disease,
Alzheimer's disease and other dementias, Parkinson's disease (PD)
and PD-related disorders, multi infarct dementia, Frontotemporal
dementia, pseudo-dementia, Prion disease, Motor neuron diseases,
Huntington's disease, spinocerebellar ataxia, and spinal muscular
atrophy.
[0065] In one embodiment, said use is in combination with another
pharmaceutically active drug or therapy, in particular radiation
therapy, chemotherapy agents, targeted drugs and immune check point
inhibitor drugs.
[0066] In a further aspect, the present invention relates to a
method of treatment of a disease selected from hyperproliferative
disorders, inflammatory disorders and/or neurodegenerative
disorders, comprising administering a compound according to the
present invention, or a composition according to the present
invention to a patient in need thereof.
[0067] The present invention also relates to the use of a compound
or composition according to the present invention as defined above,
for the manufacture of a medicament for the treatment of a disease
associated with, accompanied by, caused by Axl/Mer and CSF1R. The
present invention also relates to a method of treatment of a
disease associated with, accompanied by, caused by and/or induced
by Axl/Mer and CSF1R, said method comprising the administration of
a compound according to the present invention to a patient in need
thereof. In one embodiment, the disease associated with,
accompanied by, caused by and/or induced by Axl/Mer and CSF1R is a
disease selected from hyperproliferative disorders, inflammatory
disorders and neurodegenerative disorders, all as defined further
above.
[0068] Without wishing to be bound by any theory, the present
inventors believe that the compounds of the present invention are
efficient inhibitors of Axl/Mer and CSF1R and thus, are suitable
for the treatment of disorders associated with, accompanied by,
caused by Axl/Mer and CSF1R, in particular their hyper-function,
and thereby have an effect on one or several of cell survival,
proliferation, autophagy, vascular smooth muscle homeostasis,
migration, adhesion, angiogenesis, platelet aggregation, thrombus
stabilization, erythropoiesis, oligodendrocyte cell survival,
osteoclast function, innate immunity, inflammation, phagocytosis of
apoptotic cells and/or natural killer cell differentiation.
[0069] The compounds of the invention are capable of inhibiting
cell proliferation and thus, are suitable for the treatment and/or
prevention of Axl/Mer and CSF1R induced hyperproliferative
disorders, particularly selected from the group comprising cancer,
especially immune-suppressive cancer and refractory cancer, and
primary tumor metastases. In a preferred embodiment of the
invention, the Axl/Mer and CSF1R induced disorders are associated
with Axl/Mer and CSF1R overexpression and/or hyperactivity, e.g. an
increased degree of auto-phosphorylation compared to normal tissue.
The hyperproliferative disorder may be a cancer, preferably a
cancer selected from breast cancer, colon cancer, prostate cancer,
lung cancer, gastric cancer, ovarian cancer, endometrial cancer,
renal cancer, hepatocellular cancer, thyroid cancer, uterine
cancer, esophagus cancer, squamous cell cancer, leukemia,
osteosarcoma, melanoma, glioblastoma and neuroblastoma. In an
especially preferred embodiment, the disorders are selected from
breast cancer, glioblastoma, renal cancer, non-small cell lung
cancer (NSCLC), and melanoma.
[0070] In an especially preferred embodiment, the disorders are
selected from breast cancer, glioblastoma, renal cancer, non-small
cell lung cancer (NSCLC), and melanoma. Examples for disorders
associated with, accompanied by, caused by and/or induced by
Axl/Mer and CSF1R hyper-function are acute lymphoblastic leukemia,
acute myeloid leukemia, adrenocortical carcinoma, aids-related
cancers, aids-related lymphoma, anal cancer, appendix cancer,
astrocytomas, atypical teratoid/rhabdoid tumor, basal cell
carcinoma, bile duct cancer, bladder cancer, bone cancer,
osteosarcoma and malignant fibrous histiocytoma, brain stem glioma,
brain tumor, central nervous system atypical teratoid/rhabdoid
tumor, astrocytomas, craniopharyngioma, ependymoblastoma,
ependymoma, medulloblastoma, medulloepithelioma, pineal parenchymal
tumors of intermediate differentiation, supratentorial primitive
neuroectodermal tumors and pineoblastoma, brain and spinal cord
tumors, breast cancer, bronchial tumors, burkitt lymphoma,
carcinoid tumor, gastrointestinal cancer, central nervous system
(CNS) lymphoma, cervical cancer, chordoma, chronic lymphocytic
leukemia, chronic myelogenous leukemia, chronic myeloproliferative
disorders, colon cancer, colorectal cancer, craniopharyngioma,
cutaneous t-cell lymphoma, mycosis fungoides, sezary syndrome,
endometrial cancer, ependymoblastoma, ependymoma, esophageal
cancer, esthesioneuroblastoma, ewing sarcoma family of tumors,
extracranial germ cell tumor, extragonadal germ cell tumor,
extrahepatic bile duct cancer, intraocular melanoma,
retinoblastoma, gallbladder cancer, gastric (stomach) cancer,
gastrointestinal carcinoid tumor, gastrointestinal stromal tumor
(gist), gastrointestinal stromal cell tumor, extracranial germ cell
tumor, extragonadal germ cell tumor, ovarian germ cell tumor,
gestational trophoblastic tumor, glioma, hairy cell leukemia, head
and neck cancer, heart cancer, hepatocellular (liver) cancer,
histiocytosis, hodgkin lymphoma, hypopharyngeal cancer, intraocular
melanoma, islet cell tumors (endocrine pancreas), kaposi sarcoma,
renal cell cancer, kidney cancer, langerhans cell histiocytosis,
laryngeal cancer, acute lymphoblastic leukemia, acute myeloid
leukemia, chronic lymphocytic leukemia, chronic myelogenous
leukemia, leukemia, lip and oral cavity cancer, liver cancer, lung
cancer, non-small cell lung cancer, small cell lung cancer,
aids-related lymphoma, burkitt lymphoma, (cutaneous t-cell
lymphoma, hodgkin lymphoma, non-hodgkin lymphoma, primary central
nervous system lymphoma, macroglobulinemia, malignant fibrous
histiocytoma of bone and osteosarcoma, medulloblastoma,
medulloepithelioma, melanoma, melanoma intraocular (eye), merkel
cell carcinoma, mesothelioma, metastatic squamous neck cancer with
occult primary, mouth cancer, multiple endocrine neoplasia
syndromes, multiple myeloma/plasma cell neoplasm, myelodysplastic
syndromes, myelodysplastic/myeloproliferative neoplasms,
myelogenous leukemia, myeloid leukemia, myeloma (multiple),
myeloproliferative disorders, nasal cavity and paranasal sinus
cancer, nasopharyngeal cancer, neuroblastoma, non-hodgkin lymphoma,
non-small cell lung cancer, oral cancer, oral cavity cancer,
oropharyngeal cancer, osteosarcoma and malignant fibrous
histiocytoma of bone, ovarian cancer, ovarian epithelial cancer,
ovarian germ cell tumor, ovarian low malignant potential tumor,
pancreatic cancer, papillomatosis, parathyroid cancer, penile
cancer, pharyngeal cancer, pineoblastoma and supratentorial
primitive neuroectodermal tumors, pituitary tumor, plasma cell
neoplasm/multiple myeloma, pleuropulmonary blastoma, pregnancy and
breast cancer, prostate cancer, rectal cancer, renal cell (kidney)
cancer, transitional cell cancer, respiratory tract cancer,
retinoblastoma, rhabdomyosarcoma, salivary gland cancer, sarcoma,
ewing sarcoma, kaposi sarcoma, uterine sarcoma, nonmelanoma skin
cancer, melanoma skin cancer, skin carcinoma, small cell lung
cancer, small intestine cancer, soft tissue sarcoma, squamous cell
carcinoma, squamous neck cancer, stomach (gastric) cancer,
supratentorial primitive neuroectodermal tumors, t-cell lymphoma,
testicular cancer, throat cancer, thymoma and thymic carcinoma,
thyroid cancer, transitional cell cancer of the renal pelvis and
ureter, trophoblastic tumor, gestational cancer, ureter and renal
pelvis cancer, transitional cell cancer, urethral cancer, uterine
cancer, endometrial cancer, uterine sarcoma, vaginal cancer, vulvar
cancer, Waldenstrom macroglobulinemia and Wilms tumor.
[0071] The compounds of the present invention are efficient
inhibitors of Axl/Mer and CSF1R. The inventive compounds are
suitable for the use as a pharmaceutically active agent which are
suitable for the treatment of disorders associated with,
accompanied by, caused by Axl/Mer and CSF1R, in particular a
hyper-function thereof. The inventive compounds are thus suitable
for the treatment of Axl/Mer and CSF1R induced disorders.
[0072] Without wishing to be bound by any theory, the present
inventors believe that CSF1 is another chemokine that appears to be
a promising target for inhibiting TAM contribution to tumor
progression. CSF1 is a potent chemoattractant and is considered the
most important growth factor regulating the differentiation of
monocytes into macrophages. Thus, the present inventors believe
that the inhibition of CSF1R signaling in tumor-promoting TAMs
represents an attractive strategy to eliminate or repolarize these
cells and it is a possible of new immunotherapy for patients with
cancers.
[0073] Again without wishing to be bound by any theory, the present
inventors believe that CSF1/CSF1R blockade not only decreases the
number of TAMs, but also reprograms remaining TAMs to support
antigen presentation and bolster T cell activation within the tumor
microenvironment. This in-turn leads to reduced immune suppression
and elevated interferon responses, which restrain tumor progression
(Zhu Y et al., Cancer Res. 2014 Sep. 15; 74(18):5057-69).
[0074] In conclusion, dual inhibition CSF1R and Axl/Mer RTK is
expected to offer strong anti-cancer efficacy through immune
modulation by using distinct signal pathways for immune cells. The
present invention to provide compounds which have dual-inhibition
capabilities for CSF1R and Axl/Mer RTK. These can then be used as
pharmaceutically active agents, with a high relevance especially
for cancer therapy via immune system modulation.
[0075] The term "optionally substituted" as used herein is meant to
indicate that a hydrogen atom where present and attached to a
member atom within a group, or several such hydrogen atoms, may be
replaced by a suitable group, such as halogen including fluorine,
chlorine, C.sub.1-C.sub.3 alkyl, C.sub.1-C.sub.3 haloalkyl, e. g.
CH.sub.2F, CHF.sub.2, CF.sub.3, CH.sub.2CF.sub.3, methylhydroxyl,
hydroxyl, COOMe, C(O)H, COOH, alkoxy, in particular C1-C3 alkoxy,
e.g. OMe, or OCF.sub.3;
[0076] In one embodiment, the present invention also relates to
pharmaceutically acceptable salts of the compounds according to the
present invention.
[0077] The present invention also relates to combinations of
compounds in accordance with the present invention and another
anti-cancer agent together.
[0078] Combination of a compound in accordance with the present
invention with another anti-cancer agent has a better anti-cancer
effect. For example combinations with other anti-cancer agents may
restore sensitivity of cell lines which have become resistant to
other anti-cancer agents and combinations with other immune
checkpoint inhibitor as well as radiation also have a synergistic
efficacy. As examples of agents with which the compounds according
to the present invention may be combined, as a cytotoxicity drugs
(actinomycin, all-trans retinoic acid, azacitidine, azathioprine,
bleomycin, bortezomib, carboplatin, capecitabine, cisplatin,
chlorambucil, cyclophosphamide, cytarabine, daunorubicin,
docetaxel, doxifluridine, doxorubicin, epirubicin, epothilone,
etoposide, fluorouracil, gemcitabine, hydroxyurea, idarubicin,
imatinib, irinotecan, mechlorethamine, mercaptopurine,
methotrexate, mitoxantrone, oxaliplatin, paclitaxel, pemetrexed,
teniposide, tioguanine, topotecan, valrubicin, vemurafenib,
vinblastine, vincristine, vindesine, vinorelbine), targeted drugs
(bevacizumab, rituximab, ipilimumab, bortezomib, imatinib,
seliciclib, ado-trastuzumab, afatinib, aldesleukin, axitinib,
belinostat, bevacizumab, bortezomib, bosutinib, brentuximab
vedotin, cabozantinib, canakinumab, carfilzomib, ceritinib,
cetuximab, crizotinib, dabrafenib, dasatinib, erlotinib,
everolimus, gefitinib, ibritumomab tiuxetan, ibrutinib, idelalisib,
imatinib, lapatinib, lenvatinib, nilotinib, obinutuzumab,
ofatumumab, olaparib, palbociclib, panitumumab, panobinostat,
pazopanib, pertuzumab, ponatinib, ramucirumab, regorafenib,
rituxima, romidepsin, ruxolitinib, siltuximab, sipuleucel-T,
sorafenib, temsirolimus, tocilizumab, tofacitinib, tositumomab,
trametinib, trastuzumab, vandetanib, vemurafenib, vismodegib,
vorinosta, ziv-aflibercept), immune check point inhibitor drugs
(lpillmumab, nivolumab, pembrolizumab, atezolezumab, avelumab,
bevacixumab, tremelimumab).
[0079] Other combinations may also or instead include several
compounds in accordance with the present invention together. These
are also envisaged and encompassed by combinations in accordance
with the present invention.
[0080] The term "alkyl" refers to a monovalent straight, branched
or cyclic chain, saturated aliphatic hydrocarbon radical having a
number of carbon atoms in the specified range. Thus, for example,
"C.sub.1-C.sub.6 alkyl" refers to any of the hexyl alkyl and pentyl
alkyl isomers as well as n-, iso-, sec-, and t-butyl, n- and
isopropyl, cyclic propyl, ethyl and methyl.
[0081] The term "alkenyl" refers to a monovalent straight or
branched chain aliphatic hydrocarbon radical containing one
carbon-carbon double bond and having a number of carbon atoms in
the specified range. Thus, for example, "C.sub.2-C.sub.6 alkenyl"
refers to all of the hexenyl and pentenyl isomers as well as
1-butenyl, 2-butenyl, 3-butenyl, isobutenyl, 1-propenyl,
2-propenyl, and ethenyl (or vinyl).
[0082] The term "cycloalkyl", alone or in combination with any
other term, refers to a group, such as optionally substituted or
non-substituted cyclic hydrocarbon, having from three to eight
carbon atoms, unless otherwise defined. Thus, for example,
"C.sub.3-C.sub.8 cycloalkyl" refers to cyclopropyl, cyclobutyl,
cyclopentyl, cyclohexyl, cycloheptyl, and cyclooctyl.
[0083] The term "haloalkyl" refers to an alkyl group, as defined
herein that is substituted with at least one halogen. Examples of
straight or branched chained "haloalkyl" groups useful in the
present invention include, but are not limited to, methyl, ethyl,
propyl, isopropyl, n-butyl, and t-butyl substituted independently
with one or more halogens. The term "haloalkyl" should be
interpreted to include such substituents such as --CHF.sub.2,
--CF.sub.3, --CH.sub.2--CH.sub.2--F, --CH.sub.2--CF.sub.3, and the
like. More specifically, the term "C.sub.1-C.sub.4 haloalkyl" is
meant to include, but not to be limited to fluoromethyl,
difluoromethyl, trifluoromethyl, fluoroethyl, difluoroethyl,
trifluoroethyl, fluoropropyl, difluoropropyl, trifluoropropyl,
fluoroisopropyl, difluoroisopropyl, trifluoroisopropyl,
fluorobutyl, difluorobutyl, trifluorobutyl, with "butyl" including
all the butyl isomers, i. e. n-butyl, iso-butyl, sec-butyl and
tert-butyl; and the corresponding chloro-alkyls of any of the
foregoing, as well as C.sub.1-C.sub.4 alkyls in which more than one
hydrogen is substituted by a halogen (F, Cl, Br and/or I).
[0084] The term "heteroalkyl" refers to an alkyl group where one or
more carbon atoms have been replaced with a heteroatom, such as, O,
N, or S. For example, if the carbon atom of alkyl group which is
attached to the parent molecule is replaced with a heteroatom
(e.g., O, N, or S) the resulting heteroalkyl groups are,
respectively, an alkoxy group (e.g., --OCH.sub.3, etc.), an amine
(e.g., --NHCH.sub.3, --N(CH.sub.3).sub.2, etc.), or thioalkyl group
(e.g., --SCH.sub.3, etc.). If a non-terminal carbon atom of the
alkyl group which is not attached to the parent molecule is
replaced with a heteroatom (e.g., O, N, or S) and the resulting
heteroalkyl groups are, respectively, an alkyl ether (e.g.,
--CH.sub.2CH.sub.2--O--CH.sub.3, etc.), alkyl amine (e.g.,
--CH.sub.2NHCH.sub.3, --CH.sub.2N(CH.sub.3).sub.2, etc.), or
thioalkyl ether (e.g., --CH.sub.2--S--CH.sub.3).
[0085] The term "halogen" refers to fluorine, chlorine, bromine, or
iodine.
[0086] The term "phenyl" as used herein is meant to indicate an
optionally substituted or non-substituted phenyl group.
[0087] The term "benzyl" as used herein is meant to indicate an
optionally substituted or non-substituted benzyl group.
[0088] The term "heteroaryl" refers to (i) optionally substituted
5- and 6-membered heteroaromatic rings and (ii) optionally
substituted 9- and 10-membered bicyclic, fused ring systems in
which at least one ring is aromatic, wherein the heteroaromatic
ring or the bicyclic, fused ring system contains from 1 to 4
heteroatoms independently selected from N, O, and S, where each N
is optionally in the form of an oxide and each S in a ring which is
not aromatic is optionally S(O) or S(O).sub.2. Suitable 5- and
6-membered heteroaromatic rings include, for example, pyridyl,
pyrrolyl, pyrazinyl, pyrimidinyl, pyridazinyl, triazinyl, thienyl,
furanyl, imidazolyl, pyrazolyl, triazolyl, tetrazolyl, oxazolyl,
isooxazolyl, oxadiazolyl, thiazolyl, isothiazolyl, and
thiadiazolyl. Suitable 9- and 10-membered heterobicyclic, fused
ring systems include, for example, benzofuranyl, indolyl,
indazolyl, naphthyridinyl, isobenzofuranyl, benzopiperidinyl,
benzisoxazolyl, benzoxazolyl, chromenyl, quinolinyl, isoquinolinyl,
cinnolinyl, quinazolinyl, tetrahydroquinolinyl,
tetrahydroisoquinolinyl, isoindolyl, benzodioxolyl, benzo furanyl,
imidazo[1,2-a]pyridinyl, benzotriazolyl, dihydroindolyl,
dihydroisoindolyl, indazolyl, indolinyl, isoindolinyl,
quinoxalinyl, quinazolinyl, 2,3-dihydrobenzo furanyl, and
2,3-dihydrobenzo-1,4-dioxinyl.
[0089] The term "heterocyclyl" or "heterocycloalkyl", in particular
"C.sub.3-C.sub.10 heterocycloalkyl" refers to (i) optionally
substituted 3- to 10-membered, saturated and unsaturated but
non-aromatic monocyclic rings containing at least one carbon atom
and from 1 to 4 heteroatoms, (ii) optionally substituted bicyclic
ring systems containing from 1 to 6 heteroatoms, and (iii)
optionally substituted tricyclic ring systems, wherein each ring in
(ii) or (iii) is independent of fused to, or bridged with the other
ring or rings and each ring is saturated or unsaturated but
nonaromatic, and wherein each heteroatom in (i), (ii), and (iii) is
independently selected from N, O, and S, wherein each N is
optionally in the form of an oxide and each S is optionally
oxidized to S(O) or S(O).sub.2. Suitable 3- to 10-membered
saturated heterocycloalkyls include, for example, azetidinyl,
piperidinyl, morpholinyl, thiomorpholinyl, thiazolidinyl,
isothiazolidinyl, oxazolidinyl, isoxazolidinyl, pyrrolidinyl,
imidazolidinyl, piperazinyl, tetrahydrofuranyl, tetrahydrothienyl,
pyrazolidinyl, hexahydropyrimidinyl, thiazinanyl, thiazepanyl,
azepanyl, diazepanyl, tetrahydropyranyl, tetrahydrothiopyranyl,
dioxanyl, and azacyclooctyl. Suitable unsaturated heterocyclic
rings include those corresponding to the saturated heterocyclic
rings listed in the above sentence in which a single bond is
replaced with a double bond. It is understood that the specific
rings and ring systems suitable for use in the present invention
are not limited to those listed in this and the preceding
paragraphs. These rings and ring systems are merely
representative.
Pharmaceutical Compositions
Pharmaceutically Acceptable Salts
[0090] Examples of pharmaceutically acceptable addition salts
include, without limitation, the non-toxic inorganic and organic
acid addition salts such as the acetate derived from acetic acid,
the aconate derived from aconitic acid, the ascorbate derived from
ascorbic acid, the benzenesulfonate derived from benzensulfonic
acid, the benzoate derived from benzoic acid, the cinnamate derived
from cinnamic acid, the citrate derived from citric acid, the
embonate derived from embonic acid, the enantate derived from
enanthic acid, the formate derived from formic acid, the fumarate
derived from fumaric acid, the glutamate derived from glutamic
acid, the glycolate derived from glycolic acid, the hydrochloride
derived from hydrochloric acid, the hydrobromide derived from
hydrobromic acid, the lactate derived from lactic acid, the maleate
derived from maleic acid, the malonate derived from malonic acid,
the mandelate derived from mandelic acid, the methanesulfonate
derived from methane sulphonic acid, the naphthalene-2-sulphonate
derived from naphtalene-2-sulphonic acid, the nitrate derived from
nitric acid, the perchlorate derived from perchloric acid, the
phosphate derived from phosphoric acid, the phthalate derived from
phthalic acid, the salicylate derived from salicylic acid, the
sorbate derived from sorbic acid, the stearate derived from stearic
acid, the succinate derived from succinic acid, the sulphate
derived from sulphuric acid, the tartrate derived from tartaric
acid, the toluene-p-sulphonate derived from p-toluene sulphonic
acid, and the like. Such salts may be formed by procedures well
known and described in the art.
[0091] Other acids such as oxalic acid, which may not be considered
pharmaceutically acceptable, may be useful in the preparation of
salts useful as intermediates in obtaining a chemical compound of
the invention and its pharmaceutically acceptable acid addition
salt.
[0092] In another embodiment, the compounds of the invention are
used in their respective free base form according to the present
invention.
[0093] Metal salts of a chemical compound of the invention include
alkali metal salts, such as the sodium salt of a chemical compound
of the invention containing a carboxy group.
[0094] The chemical compounds of the invention may be provided in
unsolvated or solvated forms together with a pharmaceutically
acceptable solvent(s) such as water, ethanol, and the like.
Solvated forms may also include hydrated forms such as the
monohydrate, the dihydrate, the hemihydrate, the trihydrate, the
tetrahydrate, and the like. In general, solvated forms are
considered equivalent to unsolvated forms for the purposes of this
invention.
Administration and Formulation
[0095] The production of medicaments containing the compounds of
the invention, its active metabolites or isomers and salts
according to the invention and their application can be performed
according to well-known pharmaceutical methods.
[0096] While the compounds of the invention, useable according to
the invention for use in therapy, may be administered in the form
of the raw chemical compound, it is preferred to introduce the
active ingredient, optionally in the form of a physiologically
acceptable salt in a pharmaceutical composition together with one
or more adjuvants, excipients, carriers, buffers, diluents, and/or
other customary pharmaceutical auxiliaries. Such salts of the
compounds of the invention may be anhydrous or solvated.
[0097] In a preferred embodiment, the invention provides
medicaments comprising a compound useable according to the
invention, or a pharmaceutically acceptable salt or derivative
thereof, together with one or more pharmaceutically acceptable
carriers therefor, and, optionally, other therapeutic and/or
prophylactic ingredients. The carrier(s) must be "acceptable" in
the sense of being compatible with the other ingredients of the
formulation and not harmful to the recipient thereof.
[0098] A medicament of the invention may be those suitable for
oral, rectal, bronchial, nasal, topical, buccal, sub-lingual,
transdermal, vaginal or parenteral (including cutaneous,
subcutaneous, intramuscular, intraperitoneal, intravenous,
intraarterial, intracerebral, intraocular injection or infusion)
administration, or those in a form suitable for administration by
inhalation or insufflation, including powders and liquid aerosol
administration, or by sustained release systems. Suitable examples
of sustained release systems include semipermeable matrices of
solid hydrophobic polymers containing the compound of the
invention, which matrices may be in form of shaped articles, e.g.
films or microcapsules.
[0099] The compounds useable according to the invention, together
with a conventional adjuvant, carrier, or diluent, may thus be
placed into the form of medicament and unit dosages thereof. Such
forms include solids, and in particular tablets, filled capsules,
powder and pellet forms, and liquids, in particular aqueous or
non-aqueous solutions, suspensions, emulsions, elixirs, and
capsules filled with the same, all for oral use, suppositories for
rectal administration, and sterile injectable solutions for
parenteral use. Such medicament and unit dosage forms thereof may
comprise conventional ingredients in conventional proportions, with
or without additional active compounds or principles, and such unit
dosage forms may contain any suitable effective amount of the
active ingredient commensurate with the intended daily dosage range
to be employed.
[0100] The compounds useable according to the invention can be
administered in a wide variety of oral and parenteral dosage forms.
It will be obvious to those skilled in the art that the following
dosage forms may comprise, as the active component, either a
compound(s) useable according to the invention or a
pharmaceutically acceptable salt of a compound(s) useable according
to the invention.
[0101] For preparing a medicament from a compound useable according
to the invention, pharmaceutically acceptable carriers can be
either solid or liquid. Solid form preparations include powders,
tablets, pills, capsules, cachets, suppositories, and dispersible
granules. A solid carrier can be one or more substances which may
also act as diluents, flavouring agents, solubilizers, lubricants,
suspending agents, binders, preservatives, tablet disintegrating
agents, or an encapsulating material.
[0102] In powders, the carrier is a finely divided solid which is
in a mixture with the finely divided active component. In tablets,
the active component is mixed with the carrier having the necessary
binding capacity in suitable proportions and compacted in the shape
and size desired. Suitable carriers are magnesium carbonate,
magnesium stearate, talc, sugar, lactose, pectin, dextrin, starch,
gelatin, tragacanth, methylcellulose, sodium
carboxymethylcellulose, a low melting wax, cocoa butter, and the
like. The term "preparation" is intended to include the formulation
of the active compound with encapsulating material as carrier
providing a capsule in which the active component, with or without
carriers, is surrounded by a carrier, which is thus in association
with it. Similarly, cachets and lozenges are included. Tablets,
powders, capsules, pills, cachets, and lozenges can be used as
solid forms suitable for oral administration.
[0103] For preparing suppositories, a low melting wax, such as a
mixture of fatty acid glyceride or cocoa butter, is first melted
and the active component is dispersed homogeneously therein, as by
stirring. The molten homogenous mixture is then poured into
convenient sized moulds, allowed to cool, and thereby to solidify.
Compositions suitable for vaginal administration may be presented
as pessaries, tampons, creams, gels, pastes, foams or sprays
containing in addition to the active ingredient such carriers as
are known in the art to be appropriate. Liquid preparations include
solutions, suspensions, and emulsions, for example, water or
water-propylene glycol solutions. For example, parenteral injection
liquid preparations can be formulated as solutions in aqueous
polyethylene glycol solution.
[0104] The chemical compounds according to the present invention
may thus be formulated for parenteral administration (e.g. by
injection, for example bolus injection or continuous infusion) and
may be presented in unit dose form in ampoules, pre-filled
syringes, small volume infusion or in multi-dose containers with an
added preservative. The compositions may take such forms as
suspensions, solutions, or emulsions in oily or aqueous vehicles,
and may contain formulation agents such as suspending, stabilising
and/or dispersing agents. Alternatively, the active ingredient may
be in powder form, obtained by aseptic isolation of sterile solid
or by lyophilization from solution, for constitution with a
suitable vehicle, e.g. sterile, pyrogen-free water, before use.
[0105] Aqueous solutions suitable for oral use can be prepared by
dissolving the active component in water and adding suitable
colorants, flavours, stabilising and thickening agents, as desired.
Aqueous suspensions suitable for oral use can be made by dispersing
the finely divided active component in water with viscous material,
such as natural or synthetic gums, resins, methylcellulose, sodium
carboxymethylcellulose, or other well known suspending agents.
[0106] Also included are solid form preparations which are intended
to be converted, shortly before use, to liquid form preparations
for oral administration. Such liquid forms include solutions,
suspensions, and emulsions. These preparations may contain, in
addition to the active component, colorants, flavours, stabilisers,
buffers, artificial and natural sweeteners, dispersants,
thickeners, solubilizing agents, and the like.
[0107] In one embodiment of the present invention, the medicament
is applied topically or systemically or via a combination of the
two routes.
[0108] For administration, the compounds of the present invention
may, in one embodiment, be administered in a formulation containing
0.001% to 70% per weight of the compound, preferably between 0.01%
to 70% per weight of the compound, even more preferred between 0.1%
and 70% per weight of the compound. In one embodiment, a suitable
amount of compound administered is in the range of from 0.01 mg/kg
body weight to 1 g/kg body weight.
[0109] Compositions suitable for administration also include
lozenges comprising the active agent in a flavoured base, usually
sucrose and acacia or tragacanth; pastilles comprising the active
ingredient in an inert base such as gelatin and glycerol or sucrose
and acacia; and mouthwashes comprising the active ingredient in a
suitable liquid carrier.
[0110] Solutions or suspensions are applied directly to the nasal
cavity by conventional means, for example with a dropper, pipette
or spray. The compositions may be provided in single or multi-dose
form. In the latter case of a dropper or pipette, this may be
achieved by the patient administering an appropriate, predetermined
volume of the solution or suspension. In the case of a spray, this
may be achieved for example by means of a metering atomising spray
pump.
[0111] Administration to the respiratory tract may also be achieved
by means of an aerosol formulation in which the active ingredient
is provided in a pressurised pack with a suitable propellant such
as a chlorofluorocarbon (CFC) for example dichlorodifluoromethane,
trichlorofluoromethane, or dichlorotetrafluoroethane, carbon
dioxide, or other suitable gas. The aerosol may conveniently also
contain a surfactant such as lecithin. The dose of drug may be
controlled by provision of a metered valve.
[0112] Alternatively the active ingredients may be provided in the
form of a dry powder, for example a powder mix of the compound in a
suitable powder base such as lactose, starch, starch derivatives
such as hydroxypropylmethyl cellulose and polyvinylpyrrolidone
(PVP). Conveniently the powder carrier will form a gel in the nasal
cavity. The powder composition may be presented in unit dose form
for example in capsules or cartridges of, e.g., gelatin, or blister
packs from which the powder may be administered by means of an
inhaler.
[0113] In compositions intended for administration to the
respiratory tract, including intranasal compositions, the compound
will generally have a small particle size for example of the order
of 5 microns or less. Such a particle size may be obtained by means
known in the art, for example by micronization.
[0114] When desired, compositions adapted to give sustained release
of the active ingredient may be employed.
[0115] The pharmaceutical preparations are preferably in unit
dosage forms. In such form, the preparation is subdivided into unit
doses containing appropriate quantities of the active component.
The unit dosage form can be a packaged preparation, the package
containing discrete quantities of preparation, such as packaged
tablets, capsules, and powders in vials or ampoules. Also, the unit
dosage form can be a capsule, tablet, cachet, or lozenge itself, or
it can be the appropriate number of any of these in packaged form.
Tablets or capsules for oral administration and liquids for
intravenous administration and continuous infusion are preferred
compositions.
[0116] Further details on techniques for formulation and
administration may be found in the latest edition of Remington's
Pharmaceutical Sciences (Maack Publishing Co. Easton, Pa.).
[0117] The invention is further illustrated by the following
figures, tables and Examples which are given to illustrate the
invention, not to limit it.
Tables
[0118] Reference is now made to the figures and tables, wherein
[0119] Table 1 shows activity data in Axl, Mer and CSF1R binding
assay for selected compounds of the invention. Inhibition is
indicated as Kd with the following key: A=Kd less than 0.1 uM; B=Kd
greater than 0.1 uM, but less than 0.5 uM; C=Kd greater than 0.5
uM. "n.d."=not determined
[0120] Table 2 shows activity data of a CSF1R binding assay for
selected compounds of the invention Inhibition is indicated as
percent inhibition in percentage figures ("Percent (%)") and with
the following additional key, classifying the inhibition figures
into different classes ("Range"): A=.gtoreq.80% inhibition; 80%
inhibition>B.gtoreq.50% inhibition; 50% inhibition>C;
[0121] Table 3 shows activity data in a cellular Axl Elisa assay
(H1299) for selected compounds of the invention. Inhibition is
indicated as IC.sub.50 with the following key: A=IC.sub.50 less
than 0.5 uM; B=IC.sub.50 greater than or equal to 0.5 uM, but less
than 1.0 uM; C=IC.sub.50 greater than or equal to 1.0 uM.
[0122] Table 4 shows activity data in a cellular CSF1R Elisa assay
(THP-1) for selected compounds of the invention. Inhibition is
indicated as IC.sub.50 with the following key: A=IC.sub.50 less
than 0.5 uM; B=IC.sub.50 greater than or equal to 0.5 uM, but less
than 1.0 uM; C=IC.sub.50 greater than or equal to 1.0 uM.
[0123] Table 5 shows CSF1R activity data in a cellular M-NFS-60
viability assay; CSF1R inhibition is indicated as IC.sub.50 with
the following key: A: IC.sub.50<1.0 .mu.M; B: 1.0
.mu.M.ltoreq.IC.sub.50<10 .mu.M; C: IC.sub.50.gtoreq.10
.mu.M.
[0124] Table 6 shows Axl, Mer and CSF1R activity data in a cellular
BaF.sub.3 viability assay; inhibition is indicated as IC.sub.50
with the following key: A: IC.sub.50<1.0 .mu.M; B: 1.0
.mu.M.ltoreq.IC.sub.50<10 .mu.M; C: IC.sub.50.gtoreq.10
.mu.M.
[0125] Table 7 shows comparison data of selected compounds of the
present invention (marked with two asterisks**) vs. selected
compounds of WO 2016/166250 (marked with a single asterisk*) in
terms of binding activity (Axl and Mer and cellular activity (Axl
H1299 Elisa assay).
[0126] Table 8 Summarizes compounds 1-179 in terms of their
structures and corresponding characteristics.
[0127] FIGS. 1 and 2 show comparison data of compound 4 of the
present invention (marked with two asterisks** and diamond) vs.
compound 27 of WO2016/166250 (marked with single asterisk* and open
square) in mouse models EMT-6 and MV4-11; EMT-6 measures triple
inhibition of Axl, Mer and CSF1R. MV4-11 measures inhibition of
Axl.
EXAMPLES
[0128] The invention is now further described by reference to the
following examples which are intended to illustrate, not to limit
the scope of the invention.
Example 1 Kinase Binding Assay for Axl and Mer
Binding Assay Principle
[0129] LanthaScreen.RTM. Eu Kinase Binding assays were conducted at
Life Technologies using the manufacturer's specifications for each
kinase indicated.
[0130] Briefly, the principle behind this assay is based upon the
binding and displacement of an Alexa Fluor 647-labeled tracer to
the kinase of interest. Binding of the tracer to the kinase is
detected using an EU-labeled anti-tag antibody. Simultaneous
binding of both the tracer and antibody to the kinase gives rise to
a FRET-signal. Binding of an inhibitor to the kinase competes for
binding with the tracer, resulting in a loss of FRET-signal.
Binding Assay Protocol for AXL and Mer
[0131] A given compound was diluted in DMSO to make a stock
compound solution. The stock compound solution was serially diluted
over eight steps in DMSO. Each diluted compound solution in DMSO
was diluted in kinase buffer. Afterwards four kinds of working
solution were prepared. First, Tracer-Working solution consists of
Tracer 236 and Kinase Buffer. Second, Axl, Mer, Tyro3 or
Met/anti-GST-AB-Working solution contained one of the kinases Axl,
and Mer or anti-GST-AB (=anti glutathione-S-transferase antibody)
in Kinase Buffer. Third, anti-GST-AB-Working solution was made with
anti-GST-AB and Kinase Buffer. Last, in the DMSO-Working solution
DMSO was added to Kinase Buffer to a final concentration to 3%.
Each of the four kinds of working solutions were separately added
to the assay plate and then incubated for 1 h at room temperature.
After incubation, the assay plate was measured with respect to the
FRET-Signal with the EnVision (Perkin Elmer) using the program
LanthaHTRF-Assay. Data evaluation was done in the Quattro Workflow
software. Kd (the equilibrium dissociation constant) values were
calculated relative to vehicle (DMSO) control wells.
[0132] Table 1 summarizes the results obtained for AXL and Mer
kinases binding assays
Example 2: Kinase Binding Assay for CSF1R
Binding Assay Principle
[0133] KINOMEscan.TM. is based on a competition binding assay that
quantitatively measures the ability of a compound to compete with
an immobilized, active-site directed ligand. The assay is performed
by combining three components: DNA-tagged kinase; immobilized
ligand; and a test compound. The ability of the test compound to
compete with the immobilized ligand is measured via quantitative
PCR of the DNA tag.
Binding Assay Protocol for CSF1R
[0134] E. coli were grown to log-phase and infected with T7 phage
and incubated with shaking at 32.degree. C. until lysis. The
lysates were centrifuged and filtered to remove cell debris. The
remaining kinases were produced in HEK-293 cells and subsequently
tagged with DNA for qPCR detection. Streptavidin-coated magnetic
beads were treated with biotinylated small molecule ligands for 30
minutes at room temperature to generate affinity resins for kinase
assays. The liganded beads were blocked with excess biotin and
washed with blocking buffer (SeaBlock (Pierce), 1% BSA, 0.05% Tween
20, 1 mM DTT) to remove unbound ligand and to reduce non-specific
binding. Binding reactions were assembled by combining kinases,
liganded affinity beads, and test compounds in 1.times. binding
buffer (20% SeaBlock, 0.17.times.PBS, 0.05% Tween 20, 6 mM DTT).
Test compounds were prepared as 111.times. stocks in 100% DMSO. Kds
were determined using an 11-point 3-fold compound dilution series
with three DMSO control points. All compounds for Kd measurements
are distributed by acoustic transfer (non-contact dispensing) in
100% DMSO. The compounds were then diluted directly into the assays
such that the final concentration of DMSO was 0.9%. In case of %
inhibition, test compounds were prepared as 40.times. stocks in
100% DMSO and directly diluted into the assay. All reactions
performed in polypropylene 384-well plate. Each was a final volume
of 0.02 ml. The assay plates were incubated at room temperature
with shaking for 1 hour and the affinity beads were washed with
wash buffer (1.times.PBS, 0.05% Tween 20). The beads were then
re-suspended in elution buffer (1.times.PBS, 0.05% Tween 20, 0.5
.mu.M non-biotinylated affinity ligand) and incubated at room
temperature with shaking for 30 minutes. The kinase concentration
in the eluates was measured by qPCR.
[0135] The CSF1R kinase binding activity results are shown in
tables 1 (K.sub.d) and 2 (percent inhibition at a concentration of
0.1 uM of compound to be tested).
Data Analysis
[0136] Binding constants (Kds) were calculated with a standard
dose-response curve using the Hill equation:
Response = Background + Signal - Background 1 + ( KD Hill Slope /
Dose Hill Slope ) ##EQU00001##
[0137] The Hill Slope was set to -1.
[0138] Curves were fitted using a non-linear least square fit with
the Levenberg-Marquardt algorithm.
[0139] Binding % inhibition were calculated with as below:
% inhibition = 100 - [ test compound signal - positive control
signal negative control signal - positive control signal ] .times.
100 ##EQU00002## [0140] negative control=DMSO (100% Ctrl) [0141]
positive control=control compound (0% Ctrl)
Example 3: Cellular Axl Inhibition Assay
Axl Cellular Assay Principle
[0142] Axl is expressed in several of malignant cancer cells;
especially H1299 cell endogenously expresses Axl; and an increased
phosphorylation of Axl (p-Axl) can be observed, when induced by the
Axl ligand, Ga6.
Enzyme-Linked Immunosorbent Assay (ELISA) Using H1299 Cells
[0143] The H1299 cells should be 40-80% confluent in the cell
culture flask. Then cells are washed with DPBS and trypsinized.
After centrifugation, the cell pellet is re-suspended with
pre-warmed culture medium (RPMI1640, 10% FBS) and cells are
separated by pipetting 6 times up and down with a 10 ml serological
pipette. The cell suspension was diluted to a final concentration
of 1.25.times.10.sup.5 cells/ml. Then 80 .mu.l/well cell suspension
is added to the plate. Cell-plates are incubated at 37.degree.
C./5% CO.sub.2 overnight. Compounds were dissolved by DMSO to 10
mM, and kept in nitrogen cabinet. Prepare the compounds stock
plate, the reference compounds concentration starts at 0.50 mM (5
.mu.l+95 .mu.l DMSO), and the tested compounds start at 1.5 mM (6
.mu.l+34 .mu.l DMSO); Then make a 3-fold dilution series (10 doses)
compounds. Then dilute compounds by the medium in the mid plate (2
.mu.l compounds+198 .mu.l medium). Transfer 20 .mu.l diluted
compounds from mid plate to the cell coated plate. Cell-plates are
incubated at 37.degree. C./5% CO.sub.2 for 24 h. Add 25 .mu.l of
Human Gas6 (1000 ng/ml) to 96-well plate; the final concentration
of Human Gas6 is 200 ng/ml. Incubate the plates at 37.degree. C./5%
CO.sub.2 for 1 h. Remove medium and wash cells once with 300
ul/well ice-cold 1.times.PBS. Then remove PBS and add 100
.mu.l/well ice-cold 1.times. Cell Lysis Buffer to 96-well plate.
The plate is shaken for 40 min at 4.degree. C. Then plates can be
stored at -20.degree. C. After the micro-wells strips have reached
room temperature, break off the required number of micro-wells.
Place the micro-wells in the strip holder. Unused micro-wells must
be resealed and stored at 4.degree. C. immediately. Add 60 .mu.l of
cell lysate to the appropriate well. Seal with tape and press
firmly onto top of micro-wells. Incubate the plate at 37.degree. C.
for 2 h. Gently remove the tape and wash wells 5 times with
1.times. Wash Buffer, 300 .mu.l each time for each well. Add 100
.mu.l of reconstituted Detection Antibody (green color) to each
well. Seal with tape and incubate the plate at 37.degree. C. for 1
h. Repeat wash procedure. Add 100 .mu.l of reconstituted HRP Linked
secondary antibody to each well. Seal with tape and incubate the
plate at 37.degree. C. for 30 min. Repeat wash procedure. Add 100
.mu.l of TMB Substrate to each well. Seal with tape and incubate
the plate at 37.degree. C. for 15 min. Add 100 .mu.l of STOP
Solution to each well. Shake gently for a few seconds. Read
absorbance at 450 nm within 30 min after adding STOP Solution.
Calculations and Formulas: The following calculations and formulas
are used for data analysis using XL Fit software: The curve fitting
was made using the Sigmoidal Dose-Response Model (Fit model, 205).
The data are reported in table 3 (shown further below).
Example 4: Cellular CSF1R Inhibition Assay
CSF1R Cellular Assay Principle
[0144] The normal proto-oncogene c-fms encodes the macrophage
growth factor (M-CSF, also known as CSF1) receptor involved in
growth, survival, and differentiation along the monocyte-macrophage
lineage. THP-1 cells act as a model for human monocytes. It
confirms that an increased phosphorylation of CSF1R (p-CSF1R) can
be observed, when induced by CSF1R ligand, CSF1. M-NFS-60 cells are
a mouse myeloid cell line that shows CSF1-dependent proliferation,
i.e. it requires CSF1 for growth.
Enzyme-Linked Immunosorbent Assay (ELISA) Using THP-1 Cells
[0145] The THP-1 cells should be maintained from 5.times.10.sup.5
to 1.times.10.sup.6 cells/ml in the cell culture flask. After
centrifugation, the supernatant is aspirated. The cell pellet is
re-suspended with pre-warmed growth medium (RPMI1640, 10% FBS, 0.05
mM 2-Mercaptoethanol) and cells are separated by pipetting 6 times
up and down with a 10 ml serological pipette. The cell suspension
has to be diluted to a final concentration of 2.times.10.sup.5
cells per 80 .mu.l with pre-warmed growth medium. 80 .mu.l of cell
suspension is added to all wells of plates. Cell-plates are
incubated at 37.degree. C./5% CO.sub.2. Dissolve compound in 100%
DMSO to make 10 mM stock solutions (Aliquots stored in the Nitrogen
Cabinet). Dilute 10 mM compounds solutions for 2-fold in 100% DMSO
to get 5 mM solutions (10 .mu.l+10 .mu.l DMSO), and then make a
3-fold serial dilution (10 .mu.l+20 .mu.l DMSO) for 10 doses. For
reference compounds: Dilute 10 mM compounds solutions for 20-fold
in 100% DMSO to get 0.5 mM solutions (2 .mu.l+38 .mu.l DMSO), and
then make a 3-fold serial dilution (10 .mu.l+20 .mu.l DMSO) for 10
doses. Transfer 2 .mu.l diluted compounds to 198 .mu.l medium to
get solution of 50 .mu.M concentration at top point for compounds
and 5 .mu.M for reference compounds. Then transfer 20 .mu.l cpd
solution into assay plate (Duplicate wells). The plate is incubated
at 37.degree. C./5% CO.sub.2 for 24 h. The final concentration of
DMSO in all wells is 0.2%. Reconstitute human M-CSF at 50 .mu.g/ml
in sterile water. (Aliquots stored in -80.degree. C.). 1:1000
dilute human M-CSF by fresh medium to 50 ng/ml. After 24 h
compounds treatment, transfer 25 .mu.l diluted human M-CSF (50
ng/ml) to cell plate according to the plate map. The final
concentration of human M-CSF in all wells is 10 ng/ml.
[0146] The plates is incubated at 37.degree. C./5% CO.sub.2 for 5
min. In the beginning the appropriate amount of lysis buffer
complete (100 .mu.L/well) is prepared and stored on ice. After 5
min human M-CSF stimulation, wash plates once with 300 .mu.L ice
cold PBS. Then 100 .mu.l lysis buffer is added to each well. The
plate is shacked for 40 min at 4.degree. C. Plates can now be
stored at -20.degree. C. After equilibrate the micro-well strips at
room temperature, break off the required number of micro-wells.
Place the micro-wells in the strip holder. Unused micro-wells must
be resealed and stored at 4.degree. C. immediately. Add 90 .mu.l of
cell lysate to the appropriate well. Seal with tape and press
firmly onto top of micro-wells. Incubate the plate for 2 h at
37.degree. C. Gently remove the tape and wash wells 4 times with
1.times. Wash Buffer, 200 .mu.l each time for each well. Add 100
.mu.l of reconstituted Detection Antibody (green color) to each
well. Seal with tape and incubate the plate at 37.degree. C. for 1
h. Repeat wash procedure. Add 100 .mu.l of reconstituted HRP-Linked
secondary antibody to each well. Seal with tape and incubate the
plate for 30 min at 37.degree. C. Repeat wash procedure. Add 100
.mu.l of TMB Substrate to each well. Seal with tape and incubate
the plate for 10 min at 37.degree. C. or 30 min at 25.degree. C.
Calculations and Formulas: The following calculations and formulas
are used for data analysis using XL Fit software: The curve fitting
was made using the Sigmoidal Dose-Response Model (Fit model, 205).
The data are reported in table 4 (shown further below).
Cell Titer-Glo (CTG) Assay Using M-NFS-60 Cells
[0147] M-NFS-60 cells were cultured in RPMI-1640 medium containing
10% FBS, 0.05 mM 2-Mercaptoethanol, penicillin (100
U/ml)/streptomycin (100 mg/ml) and supplemented with 62 ng/ml
recombinant human M-CSF at 37.degree. C./5% CO.sub.2. Aliquots of
the cells were seeded into a 96-well plate (2.times.10.sup.5 cells
in 100 .mu.l/well) and then incubated at 37.degree. C./5% CO.sub.2
1 h. Dissolve compound in 100% DMSO to make 10 mM stock solutions
(Aliquots stored in the Nitrogen Cabinet). Dilute 10 mM compounds
solutions for 2-fold in 100% DMSO to get 5 mM solutions (10
.mu.l+10 .mu.l DMSO), and then make a 3-fold serial dilution (10
.mu.l+20 .mu.l DMSO) for 10 doses. For reference compounds: Dilute
10 mM compounds solutions for 20-fold in 100% DMSO to get 0.5 mM
solutions (2 .mu.l+38 .mu.l DMSO), and then make a 3-fold serial
dilution (10 .mu.l+20 .mu.l DMSO) for 8 doses. The 96-well plate
was equilibrated at room temperature for 30 min. 50 .mu.l of
CellTiter-Glo.RTM. Reagent added to each well and mixed for 2 min
on an orbital shaker to induce cell lysis. The 96-well plate was
incubated at room temperature for 10 min to stabilize luminescent
signal. Luminescent was measured at 700 nm by EnVison plate reader.
Calculations and Formulas: The following calculations and formulas
are used for data analysis using XL Fit software: The curve fitting
was made using the Sigmoidal Dose-Response Model (Fit model, 205).
The data are reported in table 5 (shown further below).
Example 5: Cellular Axl, Mer and CSF1R Inhibition Measured by
BaF.sub.3-Viability Assay
BaF3 Cell-Based Assay Principle
[0148] In this system, IL-3-dependent Ba/F3 cells are modified to
express an activated recombinant kinase (in the present case Axl,
Mer and CSF1R). Following removal of IL-3, the modified cells are
dependent on the activity of the recombinant kinase for survival
and proliferation. Inhibition of such kinase(s) through test
compounds reduces survival and proliferation of the cells.
BaF3 Cell-Based Assay Protocol
[0149] Cell lines were maintained in growth media (GM) consisting
of RPMI 1640 supplemented with 10% FBS. Cells in logarithmic-phase
growth were harvested and 5,000 cells were distributed into each
well of a 384-well plate in 50 .mu.l GM. Parental cells (or
experimental assays where specified) were further supplemented with
2 ng/ml IL-3 to support cell growth and survival. Fifty nanoliters
of the indicated reference standard or experimental compound were
added to appropriate wells (in duplicate) and the cells were
cultured for 48 h at 37.degree. C./5% CO.sub.2. Viability was
determined by adding 10 .mu.l Cell Titer Glo and measuring
luminescence, which is reported as relative light units (RLU)
measured in counts per second. The following calculations and
formulas are used for data analysis using GraphPad Prism software:
The curve fitting was made using the Dose response
curves-Inhibition. The data are reported in table 6 (shown further
below).
Example 6: Comparison of Selected Compounds of Present Invention
Vs. Selected Compounds of WO 2016/166250 in Terms of Binding
Activity (Axl and Mer) and Cellular Activity
[0150] Table 7 shows the superiority of compounds 4, 10, 16 and 92
of the present invention (marked with **) vs. structurally similar
compounds 27, 64, 22 and 48 respectively, of WO 2016/166250 (marked
with *) in terms of their binding activity to Axl and Mer and in
terms of their cellular activity in the H1299 Elisa assay. The data
are extracted from tables 1, 2 and 3 and are reported further below
in table 7.
Example 7: EMT-6 and MV4-11 Mouse Models
a) EMT-6 Mouse Model
Syngeneic Model
[0151] Axl/Mer has been also demonstrated to be constitutive and
inducible expressed on multiple immune cells, particularly
dendritic cells (DCs) and macrophages, which act as negative
feedback to balance the pro-inflammatory signaling. In case of
CSF1R, it mediates tumorigenesis in tumor-immune microenvironments
and is expressed on myeloid cell tumor-associated macrophages
(TAMs); elevated serum CSF1, increased numbers of TAMs in tumors,
and high expression of tissue CSF1 and/or CSF1R are associated with
poor prognosis in patients with various cancers. Mouse syngeneic
tumor models are widely used tools to demonstrate activity of novel
anti-cancer immunotherapies. The EMT-6 syngeneic model contains
significantly higher amount of myeloid cells and also a higher
percentage of TAMs. Therefore, EMT-6 is a suitable model for
immune-oncology drug efficacy effects through changes in target
immune cells.
EMT-6 Syngeneic Tumor Model Treatment Protocol
[0152] BALB/c mice (6-8 week female, Beijing Vital River Laboratory
Animal Technology Co., Ltd.) were inoculated subcutaneously at the
upper right flank with EMT-6 tumor cells (1.times.10.sup.6) in 0.1
mL of PBS for tumor development. For the study, the treatments were
started on day 6 after tumor inoculation, mice were dosed orally
with each test compounds or vehicle daily. Tumor volumes were
measured every 3 days with a digital caliper, and were calculated
using the formula: V=0.5a.times.b.sup.2 where a and b are the long
and short diameters of the tumor in mm, respectively. Body weight
was measured every 3 days. All the procedures related to animal
handling, care and the treatment in the study were performed
according to the guidelines approved by the Institutional Animal
Care and Use Committee (IACUC) of WuXi AppTec following the
guidance of the Association for Assessment and Accreditation of
Laboratory Animal Care (AAALAC). At the time of routine monitoring,
the animals were daily checked for any effects of tumor growth and
treatments on normal behavior such as mobility, food and water
consumption (by looking only), body weight gain/loss (body weights
were measured three times a week), eye/hair matting and any other
abnormal effect as stated in the protocol. Death and observed
clinical signs were recorded on the basis of the numbers of animals
within each subset. The data are reported in FIG. 1 exemplarily for
compound 4 of the present invention vs. structurally similar
compound 27 of WO 2016/166250 and demonstrate that the compounds
according to the present invention delay tumor growth to a
significantly larger extent, than other compounds or vehicle.
b) MV4-11 Mouse Model
Xenograft Model
[0153] High expression of Axl or Gas6 in acute myeloid leukemia
(AML) patients is prognostic for poor survival outcome. AML cells
induce expression and secretion of the Axl ligand Gas6 by bone
marrow-derived stromal cells (BMDSCs). Gas6 in turn mediates
proliferation, survival, and chemoresistance of Axl-expressing AML
cells. This Gas6/Axl paracrine axis between AML cells and BMDSCs
establishes a chemoprotective tumor cell niche that can be
abrogated by Axl-targeting approaches. The Axl/Gas6 axis promotes
leukemia cell proliferation and chemoresistance.
MV4-11 Xenograft Tumor Model Treatment Protocol
[0154] BALB/c nude mice (6-8 week female, Shanghai SIPPR/BK
Laboratory Animal Co., LTD.) were inoculated subcutaneously at the
upper right flank with MV4-11 tumor cells (1.times.10.sup.7) in 0.1
mL of PBS+Matrigel (1:1) for tumor development. For the study, the
treatments were started on day 10 after tumor inoculation, mice
were dosed orally with each test compounds or vehicle daily. Tumor
volumes were measured every 3 days with a digital caliper, and were
calculated using the formula: V=0.5a.times.b.sup.2 where a and b
are the long and short diameters of the tumor in mm, respectively.
Body weight was measured every 3 days. All the procedures related
to animal handling, care and the treatment in the study were
performed according to the guidelines approved by the Institutional
Animal Care and Use Committee (IACUC) of WuXi AppTec following the
guidance of the Association for Assessment and Accreditation of
Laboratory Animal Care (AAALAC). At the time of routine monitoring,
the animals were daily checked for any effects of tumor growth and
treatments on normal behavior such as mobility, food and water
consumption (by looking only), body weight gain/loss (body weights
were measured three times a week), eye/hair matting and any other
abnormal effect as stated in the protocol. Death and observed
clinical signs were recorded on the basis of the numbers of animals
within each subset. The data are reported exemplarily in FIG. 2 for
compound 4 of the present invention vs. structurally similar
compound 27 of WO 2016/166250 and demonstrate that the compounds
according to the present invention delay tumor growth to a
drastically larger extent than other compounds or vehicle.
[0155] Data for EMT-6 and MV4-11 are shown in FIGS. 1 (EMT-6
syngeneic model efficacy study) and 2 (MV4-11 xenograft model
efficacy study).
Example 8: Derivatization of the Dimethoxyquinoline General
Scaffold
[0156] The presented compounds underwent derivatization according
to the methods outlined below (Schemes 1-21). Resulting derivatives
were examined for kinase binding, cellular activity and in vivo
activity, using the assays described above (Example 1-7), and the
results are summarized in Tables 1-7 and FIGS. 1 & 2. The
synthesized compounds 1-179 are shown in Table 8, below.
##STR00187##
General Procedure for Synthesis of C1
[0157] A method to prepare compounds of C1 is shown in Scheme 1.
The reaction of A1 and B1 is carried out in the presence of the
HATU (O-(7-Azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluronium
hexafluorophosphate), TEA in a DMF to give C1. Alternatively, the
reaction of A1 and B1 carried out in the presence of the EDCI
(1-Ethyl-3-(3-dimethylaminopropyl)carbodiimide) in a pyridine to
give C1.
##STR00188##
General Procedure for Synthesis of A3
[0158] To a HCl (3 M, 180 mL) was added compound A2 (20.0 g, 53.1
mmol) slowly at 10.degree. C., then the mixture was cooled to
0.degree. C., and a solution of NaNO.sub.2 (11.8 g, 55.8 mmol) in
water (50 mL) was added, the mixture turn to a brown solution
(solution A). To a mixture of ethyl 4-chloro-3-oxo-butanoate (28.2
g, 170 mmol) and NaOAc (66.1 g, 812 mmol) in H.sub.2O (2000
mL)/EtOH (500 mL) was dropwise added solution A at 0-5.degree. C.
Then the mixture was stirred at 10.degree. C. for 1 hour. Yellow
powder precipitated out from the reaction mixture. This phenomenon
indicated that the reaction worked, and LCMS showed 80% desired MS
value. The mixture was filtrated, the filter cake was washed with
water (500 mL) and dried over high vacuum to give 35.0 g of
compound. A3 (yield: 72.2%) as a yellow powder.
General Procedure for Synthesis of A4
[0159] To a solution of compound A3 (35.0 g, 117.17 mmol) in
absolute EtOH (300 mL) was added KOAc (23.0 g, 234 mmol). The
mixture was heated to reflux at 78.3.degree. C. for 6 hours. The
reaction mixture was a yellow solution. LCMS showed 86.7% desired
MS value. The mixture was cooled to room temperature, Most EtOH was
removed under reduced pressure, the residue was partitioned between
EtOAc (800 mL) and H.sub.2O (800 mL). The aqueous was extracted
with EtOAc (800 mL). The combined organic extract was washed with
brine (900 mL.times.2), dried over anhydrous Na.sub.2SO.sub.4,
filtered and concentrated under reduced pressure to give 27.0 g of
compound A4 (yield: 87.9%) as a yellow powder.
General Procedure for Synthesis of A5
[0160] To a solution of compound A4 (10.0 g, 38.1 mmol) in DMF (100
mL) was added NaH (3.05 g, 76.3 mmol, 60% dispersion in mineral
oil) at 0.degree. C., then the mixture was stirred at 20.degree. C.
for 30 minutes, then CF.sub.3CH.sub.2OTf (17.7 g, 76.3 mmol) was
added to the mixture and the reaction mixture was warmed to
60.degree. C. for 2.5 hour. LCMS showed the reaction was completed.
The mixture was cooled to room temperature, diluted with water
(1000 mL), extracted with EtOAc (1000 mL.times.3). The combined
extract was washed with brine (1500 mL.times.2), dried over
anhydrous Na.sub.2SO.sub.4 and filtrated, then concentrated under
reduced pressure to 46.0 g (crude, containing a little DMF) of
compound A5 as a yellow gum
General Procedure for Synthesis of A6
[0161] To a mixture of compound A5 (40.0 g, 116 mmol) in MeCN (400
mL) was added CAN (191 g, 349 mmol) in H.sub.2O (200 mL) at
0.degree. C., then the mixture was allowed to warmed to 20.degree.
C. and stirred at 20.degree. C. for 17 hours. TLC (PE/EtOAc=1/2)
showed the reaction was completed. The mixture was extracted with
EtOAc (500 mL.times.3), the combined extracts was washed with
saturated aqueous NaHCO.sub.3 (800 mL.times.3), the organic phase
was dried over anhydrous Na.sub.2SO.sub.4, filtered, the filtrate
was concentrated under reduced pressure to give a residue. The
residue was purified by Combi flash (PE/EtOAc=4/1 to 2/1) to give
7.60 g (yield: 27.5%) of compound. A6 as a yellow solid.
General Procedure for Synthesis of A7
[0162] To a solution of compound A6 (9.15 g, 38.4 mmol), D1-2 (7.18
g, 42.3 mmol) in MeCN (100 mL) was added Cs.sub.2CO.sub.3 (31.3 g,
96.1 mmol). The reaction was stirred at 25.degree. C. for 17 hours
to give a yellow suspension. LCMS showed the reaction was
completed. The reaction mixture was filtered. The filter cake was
washed with EtOAc (300 mL). The filtrate was partitioned with EtOAc
(300 mL) and water (400 mL). The organic phase was washed with
brine (400 mL.times.2) and dried over anhydrous Na.sub.2SO.sub.4,
filtered and concentrated under reduced pressure to give the crude
residue. The residue was purified by Combi Flash (PE/EtOAc=1:0 to
3:1) to give 4.3 g of undesired isomer (yield: 40%) (less polar) as
a yellow oil, and 4.9 g of compound A7 (yield: 46%) (more polar) as
a yellow gum.
General Procedure for Synthesis of A8
[0163] To a solution of compound A7 (4.9 g, 17.5 mmol) in MeOH (33
mL) was added NaOH (2.10 g, 52.5 mmol) in H.sub.2O (8 mL). The
reaction was stirred at 20.degree. C. for 2 hours to give a yellow
solution. TLC showed the reaction was completed. Most MeOH was
removed under reduced pressure. The reaction mixture was adjusted
to pH=4 with aqueous HCl (1 M). The mixture was extracted with DCM
(110 mL.times.3). The organic layers were washed with brine (150
mL), dried over anhydrous Na.sub.2SO.sub.4, filtered and
concentrated under reduced pressure to give 4.41 g of compound A8
(yield: 100%) as a yellow gum.
General Procedure for Synthesis of 4
[0164] To a solution of compound A8 (4.41 g, 17.5 mmol), EDCI (5.03
g, 26.2 mmol) in pyridine (40 mL) was added compound B1-1 (5.30 g,
17.8 mmol). The reaction was stirred at 25.degree. C. for 17 hours
to give a yellow solution. LCMS showed the staring material was not
consumed completely. The reaction mixture was concentrated under
reduced pressure to give a residue. The residue was partitioned
between DCM (400 mL) and H.sub.2O (300 mL). The organic layer was
washed with H.sub.2O (200 mL.times.2), brine (200 mL.times.2),
dried over anhydrous Na.sub.2SO.sub.4, filtered and concentrated
under reduced pressure to give a yellow gum. The crude product was
purified by Combi Flash (EtOAc:PE=0:1 to 1:0) and the eluent was
concentrated under reduced pressure to give an off-white gum. The
product was dissolved in MeCN (30 mL) and H.sub.2O (33 mL), and
lyophilized to give 5.23 g of compound 7 (yield: 55%, purity:
98.5%) as a white powder.
##STR00189##
General Procedure for Synthesis A9
[0165] A mixture of compound A6 (1.00 g, 4.20 mmol),
Cs.sub.2CO.sub.3 (3.42 g, 10.5 mmol) in DMF (10 mL) was added D1-2
(524 mg, 3.36 mmol). The mixture was stirred at 15.degree. C. for 2
hours to give a yellow mixture. LCMS showed the reaction was
completed. The mixture was partitioned between EtOAc (50 mL) and
H.sub.2O (50 mL). The aqueous phase was extracted with EtOAc (50
mL). The combined organic extract was washed with brine (50
mL.times.2), dried over anhydrous Na.sub.2SO.sub.4, filtered and
concentrated under reduced pressure to give crude product as yellow
oil. The residue was purified by Combi flash (PE:EA=1:0 to 3:1) to
give A9 (240 mg, 21.5% yield, more polar, desired product) as a
yellow gum. LCMS (Rt=1.080 min) showed the desired MS value.
General Procedure for Synthesis A10
[0166] A mixture of A9 (240 mg, 0.902 mmol) in DCM (3 mL) was added
1,3-dibromo-5,5-dimethyl-imidazolidine-2,4-dione (155 mg, 0.541
mmol). The mixture was stirred at 15.degree. C. for 12 hours, LCMS
showed starting material was remained, and added Br.sub.2 (200 uL),
started at 15.degree. C. for 12 hours to give a yellow mixture. TLC
showed the reaction was completed. The mixture was partitioned
between with DCM (50 mL) and saturated sodium bicarbonate (30 mL),
dried over anhydrous Na.sub.2SO.sub.4 and filtrated, then
concentrated under reduced pressure to give A10 (300 mg, 96.4%
yield) as a yellow powder. LCMS (Rt=1.243 min) showed the desired
MS value.
General Procedure for Synthesis A11
[0167] To a mixture of A10 (300 mg, 0.869 mmol), Cs.sub.2CO.sub.3
(708 mg, 2.17 mmol) in dioxane (3 mL) and H.sub.2O (1 mL) was added
2,4,6-trimethyl-1,3,5,2,4,6-trioxatriborinane (327 mg, 2.61 mmol),
Pd(dppf)Cl.sub.2.CH.sub.2Cl.sub.2 (71 mg, 0.0869 mmol). The
reaction mixture was stirred at 80.degree. C. for 12 hours under
N.sub.2 atmosphere to give a black mixture. LCMS (Rt=1.186 min)
showed the reaction was completed. The mixture was cooled to room
temperature and partitioned between DCM (30 mL) and water (20 mL).
The aqueous phase was extracted with DCM (30 mL). The combined
organic extracts were washed with brine (50 mL.times.2),
concentrated under reduced pressure to give crude product as a
brown oil, which was purified by Combi flash (PE/EA=I/O to 3/1) to
give A11 (53 mg, 21.8% yield) as a yellow oil.
General Procedure for Synthesis A12
[0168] To a mixture of A11 (50 mg, 0.178 mmol) in EtOH (1 mL) was
added the solution of NaOH (7.14 mg, 0.178 mmol) in H.sub.2O (1
mL). The mixture was stirred at 15.degree. C. for 2 hours to give a
brown suspension. TLC showed the reaction was completed. The
mixture was concentrated under reduced pressure to remove EtOH. The
aqueous phase was diluted with water (30 mL), acidized with HCl (3
M) to pH=4-5 and lyophilized to give A12 (46 mg, crude) as a yellow
gum, which was used for next step.
General Procedure for Synthesis 1
[0169] To a mixture of A12 (45 mg, 178 mmol), HATU (92.5 mg, 0.243
mmol) in DMF (1 mL) was added B1-1 (48.2 mg, 0.162 mmol), TEA (45
uL) under N.sub.2 atmosphere. The mixture was stirred at 60.degree.
C. for 12 hours to give a yellow mixture. LCMS showed the reaction
was completed. The mixture was cooled to temperature and poured
into water (10 mL). The yellow solid precipitated out from the
mixture. The mixture was filtrated and the filter cake was washed
with water (5 mL) to give a crude product as a brown solid, which
was purified by prep-TLC (DCM/MeOH=10/1) and lyophilization to give
1 (42.9 mg, 49.3% yield, 99.1% purity) as a yellow powder.
##STR00190##
General Procedure for Synthesis A13
The 1.sup.st Batch:
[0170] To a solution of A6 (200 mg, 0.840 mmol), D1-3 (216 mg, 2.52
mmol), pyridine (79.7 mg, 1.01 mmol), and DMAP (308 mg, 2.52 mmol)
in dioxane (5 mL) was added Cu(OAc).sub.2 (229 mg, 1.26 mmol), the
mixture was stirred at 100.degree. C. for 17 hours in air. Crude
LCMS (Rt: 1.490 min, 1.636 min) showed the reaction was
completed.
The 2.sup.nd Batch:
[0171] To a solution of A6 (500 mg, 2.10 mmol), D1-3 (541 mg, 6.30
mmol), pyridine (199 mg, 2.52 mmol), and DMAP (770 mg, 6.30 mmol)
in dioxane (10 mL) was added Cu(OAc).sub.2 (572 mg, 3.15 mmol), the
mixture was stirred at 100.degree. C. for 17 hours in air. Crude
LCMS and TLC (PE/EtOAc=3/1) showed the reaction was completed.
[0172] the each batch mixture were combined and poured into water
(100 mL), extracted with EA (50 mL.times.3), the combined extracts
was washed with aqueous NH.sub.3.H.sub.2O (50 mL.times.3, 14%) and
brine (100 mL), dried over anhydrous Na.sub.2SO.sub.4, filtered,
the filtration was concentrated under reduced pressure to give a
residue. the residue was purified by Combi flash to give 510 mg of
A13 (total yield: 20%) as a yellow gum and 320 mg of A13-1 as a
white powder.
General Procedure for Synthesis A14
The 1.sup.st Batch:
[0173] To a solution of A13 (100 mg, 0.359 mmol, 1 eq) in DCM (5
mL) was added Br.sub.2 (57 mg, 0.36 mmol, 1 eq), the mixture was
stirred at 15.degree. C. for 17 hours to give a brown solution.
crude LCMS (RT: 0.977 min) showed the reaction was not completed,
then additional Br.sub.2 (104 mg) was added into above mixture, the
resulting mixture was stirred at 15.degree. C. for 24 hours to give
a brown solution, TLC (PE/EtOAc=3/1) showed the reaction was
completed.
The 2.sup.nd Batch:
[0174] To a solution of A13 (410 mg, 1.47 mmol) in DCM (5 mL) was
added Br.sub.2 (236 mg, 1.47 mmol), the mixture was stirred at
15.degree. C. for 17 hours to give a brown solution. TLC (plate 1,
PE/EtOAc=3/1) showed the reaction was not completed, then Br.sub.2
(500 mg) was added into above mixture, the mixture was stirred at
30.degree. C. for 1 hours, TLC (plate 2, PE/EtOAc=3/1) showed the
reaction was completed
[0175] Two combined batches mixture was poured into DCM (50 mL) and
washed with saturated aqueous NaHCO.sub.3 (50 mL.times.3), dried
over anhydrous Na.sub.2SO.sub.4, filtered, the filtrate was
concentrated under reduced pressure to give a residue, the residue
was purified by Combi flash (PE/EtOAc=10/1 to 2/1) to give 450 mg
(yield: 86%) of A14 as a white powder.
General Procedure for Synthesis A15
[0176] To a solution of A14 (450 mg, 1.26 mmol), E1 (1.37 g, 3.78
mmol) and LiCl (53 mg, 1.3 mmol) in DMF (5 mL) was added
Pd(dppf)Cl.sub.2 (184 mg, 0.252 mmol), the mixture purged with
N.sub.2 for three times and stirred at 100.degree. C. for 17 hours
to give a brown suspension. Crude LCMS (Rt: 1.233 min) and TLC
(PE/EtOAc=3/1) showed the reaction was completed, then the mixture
was treated with aqueous HCl (10 mL, 1M) to give a brown
suspension, TLC (PE/EtOAc=3/1) and LCMS showed the reaction was
completed. the pH value of the mixture was adjusted to 8 with
saturated aqueous NaHCO.sub.3, extracted with EtOAc (30
mL.times.3), the combined extracts was washed with brine (50 mL),
dried over anhydrous Na.sub.2SO.sub.3, filtered, the filtrate was
concentrated under reduced pressure to give a residue. The residue
was purified by Combi flash (PE/EtOAc=10/1) to give 100 mg (yield:
26%) of A15 as a yellow powder.
General Procedure for Synthesis A16
[0177] To a solution of A15 (100 mg, 0.327 mmol) in EtOH (3 mL) was
added NaOH (26 mg, 0.65 mmol) in H.sub.2O (1 mL), the mixture
stirred at 15.degree. C. for 3 hours to give a yellow powder. TLC
(EtOAc) showed the reaction was completed. The pH value of the
mixture was adjusted to 5, the mixture was concentrated under
reduced pressure to give a residue, the residue was dried in
vacuum. Then the crude compound was treated with aqueous NaOH (1M,
20 mL) to from the Na salt, then extracted with EtOAc (10
mL.times.2) to removed the impurity, the pH value of water phase
was adjusted to 5 with aqueous HCl (3M), extracted with DCM (30
mL.times.3), dried over anhydrous Na.sub.2SO.sub.4, filtered, the
filtrate was concentrated under reduced pressure to 60 mg (yield:
63%) of A16 as a white powder.
General Procedure for Synthesis 3
[0178] To a solution of A16 (60 mg, 0.21 mmol), B1-1 (61 mg, 0.21
mmol) and TEA (104 mg, 1.03 mmol) in DMF (1 mL) was added HATU (156
mg, 0.411 mmol), the mixture was stirred at 15.degree. C. for 17
hours to give a suspension. Crude LCMS and HPLC showed the reaction
worked well. The mixture was filtered, the filter cake was washed
with MeCN (2 mL) to give the product (about: 60 mg, HNMR:
containing DMF (7%). The product was lyophilized to give 36 mg
(yield: 31%) of 3 as a white powder.
##STR00191##
General Procedure for Synthesis A17
[0179] The mixture of A4 (10.0 g, 38.0 mmol) and Cs.sub.2CO.sub.3
(24.8 g, 76.3 mmol) in DMF (20 mL) was stirred at 25.degree. C. for
30 minutes. To the resulting mixture was added E2 (9.38 g, 76.3
mmol, 7.2 mL). The mixture was stirred at 25.degree. C. for 16
hours to form a brown mixture. LCMS showed the reaction was
completed. The mixture was partitioned between EtOAc (200 mL) and
H.sub.2O (200 mL). The aqueous phase was extracted with EtOAc (200
mL.times.2). The combined organic extract was washed with brine
(500 mL.times.2), dried over anhydrous Na.sub.2SO.sub.4, filtered
and concentrated under reduced pressure to give A17 (10.50 g,
yield: 90.5%) as a brown solid.
General Procedure for Synthesis A18
[0180] To a solution of A17 (10.5 g, 34.5 mmol) in MeCN (100 mL)
was added the solution of CAN (56.7 g, 104 mmol) in H.sub.2O (100
mL) at 0.degree. C. The mixture was stirred at 25.degree. C. for 16
hours to from a brown solution. LCMS showed the reaction was
completed. To the mixture was added EtOAc (500 mL) and saturated
NaHCO.sub.3 (500 mL). The mixture was filtered. The filtrate was
separated. The aqueous was extracted with EtOAc (500 mL.times.2).
The combined organic extract was washed with brine (800
mL.times.2), dried over anhydrous Na.sub.2SO.sub.4, filtered and
concentrated under reduced pressure to give a residue, which was
purified by Combi flash (PE/EtOAc=I/O to 4/1) to give A18 (2.40 g,
yield: 22.2%, LCMS: 63.2%) as dark yellow oil.
General Procedure for Synthesis A19
[0181] To a mixture of A18 (1.00 g, 5.04 mmol) and Cs.sub.2CO.sub.3
(3.29 g, 10.1 mmol) in DMF (10 mL) was added D1-2 (944 mg, 6.05
mmol, 0.48 mL). The mixture was stirred at 20.degree. C. for 4
hours to form a brown mixture. LCMS showed the reaction was
completed. The mixture was cooled to room temperature. The mixture
was partitioned between EtOAc (30 mL) and H.sub.2O (30 mL). The
aqueous phase was extracted with EtOAc (30 mL.times.2). The
combined organic extract was washed with brine (80 mL.times.3),
dried over anhydrous Na.sub.2SO.sub.4, filtered and concentrated
under reduced pressure to give a residue, which was purified by
Combi flash (eluent: PE/EtOAc=3/1) to give A19 (300 mg, yield:
26.4%, more polar) and A19-1 (450 mg, yield: 39.4%, less polar) as
yellow oil.
General Procedure for Synthesis A20
[0182] To a mixture of A19 (300 mg, 1.33 mmol) in EtOH (8 mL) and
H.sub.2O (4 mL) was added NaOH (159 mg, 3.98 mmol). The mixture was
stirred at 15.degree. C. for 2 hours. LCMS showed compound 2 was
remained. The mixture was stirred for another 1 hour to form a
yellow solution. TLC (eluent: EtOAc) showed the reaction was
completed. Most of EtOH was removed under reduced pressure. The
aqueous solution was diluted with H.sub.2O (5 mL) and adjusted
pH=3-4 by HCl (2 M, aq.), and then extracted with DCM (10
mL.times.3). The organic extract was dried over anhydrous
Na.sub.2SO.sub.4, filtered and concentrated under reduced pressure
to give A20 (230 mg, yield: 87.5%) as yellow gum.
General Procedure for Synthesis 6
[0183] To a mixture of A20 (240 mg, 1.21 mmol) and B1-1 (300 mg,
1.01 mmol) in DMF (6 mL) was added HATU (575 mg, 1.51 mmol) and TEA
(306 mg, 3.03 mmol, 0.42 mL). The mixture was stirred at 15.degree.
C. for 17 hours. LCMS showed the reaction was not completed. The
mixture was warmed up to 60.degree. C. and stirred for another 3
hours. LCMS showed the reaction was not completed. The mixture was
stirred at 60.degree. C. for another 17 hours to form a brown
mixture. LCMS showed the reaction was completed. The mixture was
partitioned between EtOAc (20 mL) and H.sub.2O (20 mL). The aqueous
phase was extracted with EtOAc (20 mL.times.2). The combined
organic extract was washed with brine (50 mL.times.3), dried over
anhydrous Na2SO4, filtered and concentrated under reduced pressure
to give a residue, which was purified by Combi flash (eluent:
PE/EtOAc=1/1 to 7/3) and lyophilization to give impure product. The
impure product was further purified by prep-TLC (eluent:
PE/EtOAc=1/9) to give 50 mg of the product, which still contain
impurity from LCMS. H NMR showed pure. Then the sample was further
purified by prep-HPLC (0.05% NH4HCO3). Most of MeCN was removed by
lyophilization to give 6 (25.4 mg, yield: 5.3%, LCMS: 100%) as a
white powder.
##STR00192## ##STR00193##
General Procedure for Synthesis A21
[0184] To a mixture of A4 (25 g, 95.3 mmol) in DMF (150 mL) was
added Cs.sub.2CO.sub.3 (77.7 g, 238 mmol), E3 (25 g, 185 mmol, 18.8
mL. The mixture was stirred at 25.degree. C. for 17 hours to give a
brown mixture. LCMS showed the reaction was not completed, and new
10 g of E3 was added into the above mixture, the mixture was
stirred at 25.degree. C. for 3 hours. LCMS (Rt=1.622 min) showed
the reaction was completed. The mixture was filtered, the filter
cake was washed with EtOAc (500 mL), the filtrate was washed with
water (400 mL), the aqueous phase was extracted with EtOAc (150
mL.times.2), the combined extracts were washed with water (500
mL.times.3), brine (500 mL), dried over anhydrous Na.sub.2SO.sub.4,
filtered and concentrated under reduced pressure to give A21 (18 g,
40.9 mmol) as brown oil. The crude product was used for next step
without purification.
General Procedure for Synthesis A22
[0185] To a mixture of A21 (18 g, 56.9 mmol) in MeCN (150 mL) was
added CAN (93.6 g, 170 mmol, 85.1 mL) in H.sub.2O (150 mL) at
0-5.degree. C. The mixture was stirred at 25.degree. C. for 17
hours to give a brown mixture. LCMS (Rt=1.244 min) showed the
reaction was completed. The mixture was partitioned between EtOAc
(300 mL) and water (300 mL), the aqueous phase was extracted with
EtOAc (150 mL.times.2), the combined extracts were washed with
saturated NaHCO.sub.3 (500 mL.times.3), brine (500 mL), dried over
anhydrous Na.sub.2SO.sub.4, filtered and concentrated under reduced
pressure to give a crude product, which was purified by Combi flash
(PE/EA=10:1 to 2:1) to give A22 (1.4 g, crude) as brown gum.
General Procedure for Synthesis A23
[0186] To a solution of A22 (1.2 g, 5.71 mmol), D1-1 (1.46 g, 8.56
mmol) in DMF (15 mL) was added Cs.sub.2CO.sub.3 (3.72 g, 11.4
mmol). The reaction was stirred under N2 atmosphere at 20.degree.
C. for 17 hours to give a yellow suspension. LCMS (Rt=1.526 min)
showed the reaction was completed. The reaction mixture was
partitioned between EtOAc (300 mL) and H.sub.2O (500 mL). The
separated aqueous phase was extracted with EtOAc (100 mL). The
combined organic layers were washed with H.sub.2O (200 mL.times.2),
brine (100 mL), dried over anhydrous Na.sub.2SO.sub.4, filtered and
concentrated under reduced pressure to give a residue. The residue
was purified by Combi flash (Petroleum ether/Ethyl acetate=1/0 to
3/1) to give 310 mg (yield: 19.4%, purity: 90%, more polar) of A23
as yellow brown oil, and 430 mg (yield: 29.8%, purity: 99.7%, less
polar) of A23-1 as yellow oil.
General Procedure for Synthesis A24
[0187] To a solution of A23 (310 mg, 1.23 mmol) in MeOH (3 mL) was
added a solution of NaOH (147 mg, 3.69 mmol) in H.sub.2O (4 mL).
The reaction was stirred at 20.degree. C. for 2 hours to give a
yellow solution. TLC showed the reaction was completed. The
reaction mixture was adjusted to pH=5 with aqueous HCl solution (1
M). The mixture was extracted with EtOAc (100 mL.times.2). The
organic layers were washed with brine (60 mL), dried over anhydrous
Na2SO4, filtered and concentrated under reduced pressure to give
300 mg (crude) of A24 as yellow oil.
General Procedure for Synthesis A25
[0188] To a solution of A24 (150 mg, 0.669 mmol), B1-1 (199 mg,
0.669 mmol) in pyridine (3 mL) was added EDCI (192 mg, 1.00 mmol)
under N.sub.2 atmosphere. The reaction was stirred under N.sub.2
atmosphere at 20.degree. C. for 17 hours to give a yellow mixture.
LCMS (Rt=1.459 min) showed the reaction was completed. The reaction
mixture was partitioned between EtOAc (100 mL) and water (100 mL).
The aqueous phase was extracted with EtOAc (60 mL.times.3). The
combined extracts was washed with 0.5 M aqueous NaOH solution (70
mL), H.sub.2O (70 mL), 0.5 M aqueous HCl solution (70 mL), H.sub.2O
(70 mL), brine (60 mL), dried over anhydrous Na.sub.2SO.sub.4,
filtered and concentrate under reduced pressure to give 170 mg
(yield: 50.5%) of A25 as a yellow gum.
General Procedure for Synthesis 15
[0189] To a solution of A25 (170 mg, 0.338 mmol) in THF (3 mL) was
added NMO (79 mg, 0.675 mmol), OsO.sub.4 (26 mg, 0.101 mmol) in
H.sub.2O (2 mL). The reaction was stirred at 20.degree. C. for 17
hours to give a yellow mixture. LCMS showed the reaction was
completed. The reaction mixture was added a solution of
Na.sub.2SO.sub.3 (400 mg) in H.sub.2O (4 mL), and stirred at
25.degree. C. for 20 minutes. After filtration, the filtrate was
partitioned between EtOAc (100 mL) and H.sub.2O (20 mL). The
aqueous layer was extracted with EtOAc (50 mL.times.2). The
combined organic layers were washed with H.sub.2O (40 mL.times.2),
brine (40 mL), dried over anhydrous Na.sub.2SO.sub.4, filtered and
concentrated under reduced pressure to give a residue. After
prep-HPLC (0.01% NH.sub.4HCO.sub.3) purification, the eluent was
lyophilized to give 47.9 mg (yield: 26.4%, purity: 100%) of 15 as a
white powder.
##STR00194##
General Procedure for Synthesis A26
[0190] To a solution of A6 (0.4 g, 1.68 mmol, 1 eq) in CH3CN (20
mL) was added Cs.sub.2CO.sub.3 (821 mg, 2.52 mmol, 1.5 eq) and D1-4
(186 mg, 2.02 mmol, 1.2 eq). The resulting mixture was stirred at
15.degree. C. for 16 hours to give yellow suspension. LCMS and TLC
(PE/EtOAc=1/1) showed the reaction was completed. The reaction
mixture was quenched by addition H.sub.2O (50 mL) and extracted
with EtOAc (50 mL.times.2). The combined organic layers were washed
with brine (10 mL.times.2), dried over Na.sub.2SO.sub.4, filtered
and concentrated under reduced pressure to give a residue. The
residue was purified by column chromatography (SiO.sub.2,
PE/EtOAc=1/1) to afford A26 (184 mg, 37.2% yield) as a white
solid.
General Procedure for Synthesis A27
[0191] To a solution of A26 (184 mg, 0.625 mmol, 1 eq) in MeOH (12
mL) and H.sub.2O (2 mL) was added NaOH (75 mg, 1.88 mmol, 3 eq).
The resulting mixture was stirred at 15.degree. C. for 2 hours to
give yellow solution. LCMS showed the reaction was completed. The
reaction mixture was concentrated under reduced pressure to give a
residue, then diluted with H.sub.2O (10 mL) and extracted with
EtOAc (10 mL). The water layers were adjust to pH=4 with 0.5 M HCl
(5 mL), then concentrated under reduced pressure to give A27 (300
mg, crude, contains much NaCl salt) as a white solid. The product
was used for next step directly without further purification.
General Procedure for Synthesis 19
[0192] To a solution of B1-1 (160 mg, 0.538 mmol, 1 eq) in pyridine
(3 mL) was added A27 (158 mg, 0.592 mmol, 1.1 eq) and EDCI (155 mg,
0.807 mmol, 1.5 eq). The resulting mixture was stirred at
15.degree. C. for 16 hours to give yellow solution. LCMS showed the
reaction was completed and desired product was formed. The reaction
mixture was concentrated under reduced pressure to give a residue.
The residue was added H.sub.2O (20 mL) and extracted with EtOAc (20
mL.times.2). The combined organic layers were washed with brine (10
mL.times.2), dried over Na.sub.2SO.sub.4, filtered and concentrated
under reduced pressure to give a residue, which was purified by
column chromatography (SiO.sub.2, PE/EtOAc=0/1) to afford 19 (185
mg, 63% yield) as a white solid.
General Procedure for Synthesis 17
[0193] To a solution of 19 (185 mg, 0.339 mmol, 1 eq) in MeOH (5
mL) was added NaBH.sub.4 (25.7 mg, 0.678 mmol, 2 eq) in one
portion, the resulting mixture was stirred at 0.degree. C. for 1
hour to give colorless solution. LCMS showed the reaction was
completed. The reaction mixture was quenched by addition H.sub.2O
(1 mL) and concentrated under reduced pressure to give a residue.
Then the residue diluted with H.sub.2O (20 mL) and extracted with
EtOAc (20 mL.times.2). The combined organic layers were washed with
brine (10 mL.times.2), dried over Na.sub.2SO.sub.4, filtered and
concentrated under reduced pressure to give a residue. The residue
was dissolved in CH.sub.3CN (2 mL) and H.sub.2O (5 mL), then
lyophilized to afford the product. HNMR and HPLC showed the product
with impurity, then the impure product was further purified by
prep-HPLC (column: Waters Xbridge 150*255 mobile phase: [water
(0.05% ammonia hydroxide v/v)-ACN]; B %: 40%-75%, 7 min). The
fraction was concentrated and lyophilized to give 17 (89.9 mg,
48.4% yield, 100% purity) as a white solid.
##STR00195##
General Procedure for Synthesis A28
[0194] A mixture of D1-5 (2.55 g, 10.1 mmol, 564 uL, 1.20 eq), A6
(2.00 g, 8.40 mmol, 1.00 eq) and Cs.sub.2CO.sub.3 (5.47 g, 16.80
mmol, 2.00 eq) in dioxane (17.0 mL) was heated to 100.degree. C.
for 12 h. LCMS (A28: RT=1.24 min) showed compound A6 was consumed
completely and one main peak with desired m/z was detected. The
mixture was concentrated and the filtrate was concentrated. The
residue was purified by column chromatography (SiO.sub.2, petroleum
ether/ethyl acetate=3/1 to 1/1) (TLC: petroleum ether/ethyl
acetate=1/1, A28: Rf=0.35). A28 (1.8 g, 4.55 mmol, 54.2% yield) was
obtained as yellow solid.
General Procedure for Synthesis A29
[0195] To a solution of compound A28 (1.80 g, 4.55 mmol, 1 eq) in
MeOH (12.6 mL) was added dropwise a solution of NaOH (364 mg, 9.11
mmol, 2.0 eq) in H.sub.2O (3.60 mL) at 15.about.25.degree. C. The
mixture was stirred at 15.about.25.degree. C. for 12 h. TLC
(petroleum ether/ethyl acetate=1/1, A28: Rf=0.35, A29: Rf=0.01)
showed compound A28 was consumed completely. The mixture was
concentrated. The pH value of the residue was adjusted to 5.about.6
with 4 M HCl (15.0 mL). The mixture was extracted with a solution
of DCM and IPA (3/1, 20 mL.times.4). The combined organic layers
were dried over anhydrous Na.sub.2SO.sub.4, filtered and
concentrated. Compound A29 (1.39 g, 3.78 mmol, 83.1% yield) was
obtained as yellow solid.
General Procedure for Synthesis A30
[0196] To a solution of compound A29 (200 mg, 544 umol, 1.00 eq)
and compound B1-1 (165 mg, 555 umol, 1.02 eq) in pyridine (5 V) was
added EDCI (156 mg, 816 umol, 1.50 eq). The mixture was stirred for
at 15.about.25.degree. C. 12 h under N.sub.2. LCMS (A30: RT=1.04
min) showed compound A29 was consumed completely and one main peak
with desired m/z was detected. The reaction mixture was
concentrated. Compound A30 (200 mg, 309 umol, 56.8% yield) was
obtained as brown oil.
General Procedure for Synthesis 63
[0197] A solution of Formula A30 (1.00 eq) in HCl/MeOH (50 mL) was
stirred at 15.degree. C. for 2 h. LCMS showed compound A30 was
consumed completely and one main peak with desired m/z was
detected. The mixture was concentrated under reduced pressure to
give a residue. The crude product was purified by reversed-phase
HPLC (0.1% HCl condition) (HPLC). Compound 63 (168 mg, 92.4% yield,
99.2% purity) was obtained as yellow solid.
##STR00196##
General Procedure for Synthesis A32
[0198] To a mixture of A31 (10 g, 63.7 mmol, 1 eq) in EtOH (150 mL)
was added SOCl.sub.2 (8.33 g, 70.0 mmol, 5.08 mL, 1.1 eq) slowly at
20.degree. C. (r.t.) and the resulting mixture was stirred at
20.degree. C. for 48 hours to afford a colorless solution. The
reaction was not monitored by TLC or LCMS as it is a general
reaction. The mixture was concentrated directly and then
re-evaporated with 100 mL of toluene to afford A32 (12.4 g, crude)
as a white solid.
General Procedure for Synthesis A33
[0199] To a mixture of compound A32 (12.4 g, 66.9 mmol, 1 eq) and
K.sub.2CO.sub.3 (23.1 g, 167 mmol, 2.5 eq) in MeCN (120 mL) was
added D1-1 (28.5 g, 167 mmol, 16.4 mL, 2.5 eq) and the resulting
mixture was heated to 60-70.degree. C. for 16 hours to afford a
white mixture. TLC showed two new spots. The mixture was
partitioned between EtOAc (100 mL) and H.sub.2O (100 mL). The
aqueous phase was extracted with EtOAc (100 mL.times.2). The
combined organic extract was dried over anhydrous Na.sub.2SO.sub.4,
filtered and concentrated under reduced pressure to give crude
product. The crude product was purified by combi flash
(PE/EtOAc=1/0 to 4/1 to 3/1) to afford compound A33 (9.5 g, 62.4%
yield) as a yellow oil.
General Procedure for Synthesis A34
[0200] To a solution of A33 (2 g, 8.8 mmol, 1 eq) in MeOH (20 mL)
was added wet Pd/C (200 mg, 10% purity) under N.sub.2 atmosphere.
The suspension was degassed under vacuum and purged with H.sub.2
several times. The mixture was stirred under H2 (15 psi) at
15.degree. C. for 3 hours to give black suspension. LCMS showed the
reaction was completed. The reaction mixture was filtered over a
pad of Celite and the filtrate was concentrated to obtain A34 (1.55
g, 88.8% yield, 99.4% purity) as a yellow oil.
General Procedure for Synthesis A35
[0201] To a solution of A34 (200 mg, 1.01 mmol, 1 eq) in CH.sub.3CN
(10 mL) was added 2,2,2-trifluoroethyl trifluoromethanesulfonate
(353 mg, 1.52 mmol, 1.5 eq), K.sub.2CO.sub.3 (308 mg, 2.23 mmol,
2.2 eq) and Et.sub.3N (154 mg, 1.52 mmol, 0.2 mL, 1.5 eq). The
resulting mixture was heated at 90.degree. C. (oil bath) and
stirred for 4 hours to give yellow suspension. LCMS showed the
reaction was completed, TLC (PE:EtOAc=3:1) showed part of the
starting material remain, one new spot was formed. The reaction
mixture was quenched by addition H.sub.2O (30 mL) and extracted
with EtOAc (30 mL.times.2). The combined organic layers were washed
with brine (10 mL.times.2), dried over Na.sub.2SO.sub.4, filtered
and concentrated under reduced pressure to give a residue. The
residue was purified by column chromatography (SiO.sub.2,
PE:EtOAc=3:1) to give compound A35 (110 mg, 38.9% yield) as a white
solid.
General Procedure for Synthesis A36
[0202] To a solution of A35 (110 mg, 0.394 mmol, 1 eq) in MeOH (8
mL) and H.sub.2O (2 mL) was added NaOH (78.8 mg, 1.97 mmol, 5 eq).
The resulting mixture was stirred at 15.degree. C. for 2 hours to
give colorless solution. LCMS showed the reaction was completed.
The reaction mixture was adjust to pH.about.4 with 0.5 M HCl and
stirred for 30 min, then concentrated under reduced pressure to
give A36 (121 mg, crude) as a white solid.
General Procedure for Synthesis 23
[0203] To a solution of B1-1 (95 mg, 0.32 mmol, 1 eq) and A36 (88.3
mg, 0.35 mmol, 1.1 eq) in pyridine (2 mL) was added EDCI (92 mg,
0.48 mmol, 1.5 eq). The resulting mixture was stirred at 15.degree.
C. for 16 hours to give yellow suspension. LCMS showed most of the
starting material was consumed and desired product was formed. The
reaction mixture was concentrated under reduced pressure to give a
residue, and then diluted with H.sub.2O (20 mL), extracted with
EtOAc (20 mL.times.2). The combined organic layers were washed with
brine (10 mL.times.2), dried over Na.sub.2SO.sub.4, filtered and
concentrated under reduced pressure to give a residue. The residue
was purified by column chromatography (SiO.sub.2, PE:EtOAc=1:1) to
give product, which was dissolved in CH.sub.3CN (2 mL) and H.sub.2O
(3 mL) and lyophilized to give 23 (56.5 mg, 33.3% yield, 100%
purity) as a white powder.
##STR00197##
General Procedure for Synthesis A37
[0204] To a mixture of A32 (5.45 g, 29.4 mmol, 1 eq) and
3,4-dihydro-2H-pyran (7.43 g, 88.3 mmol, 8.07 mL, 3 eq) in THF (40
mL) was added TsOH (507 mg, 2.94 mmol, 0.1 eq). The resulting
mixture was stirred at 80.degree. C. for 16 hours to give a yellow
solution. TLC (PE/EtOAc=3/1) showed the reaction was completed. The
reaction mixture was quenched with H.sub.2O (100 mL) and extracted
with EtOAc (50 mL.times.2). The organic layer were washed with
brine (20 mL.times.3), dried over anhydrous Na.sub.2SO.sub.4,
filtered and concentrated under reduced pressure to give a residue.
The residue was purified by silica gel column chromatography
(SiO.sub.2, PE/EtOAc=3:1) to give 7.01 g (yield: 88.4%) of A37 as a
yellow oil.
General Procedure for Synthesis A38
[0205] To a solution of A37 (7.01 g, 26.0 mmol, 1 eq) in MeOH (70
mL) was added wet Pd/C (1.5 g, 10% purity, 50% in water) under
N.sub.2. The suspension was degassed under vacuum and purged with
H.sub.2 several times. The mixture was stirred under H.sub.2 (15
psi) at 25.degree. C. for 16 hours to give a black suspension. TLC
(PE/EtOAc=2/1) showed the reaction was completed. The reaction
mixture was filtered by a pad of celite and the filtrate was
concentrated to give 5.84 g (yield: 93.7%) of A38 as a purple
oil.
General Procedure for Synthesis A39
[0206] To a mixture of A38 (4.20 g, 17.6 mmol, 1 eq) in DCE (25
mL), was added tetrahydropyran-4-carbaldehyde (2.20 g, 19.3 mmol,
1.1 eq), NaBH(OAc).sub.3 (9.30 g, 43.9 mmol, 2.5 eq) and HOAc (1.05
g, 17.6 mmol, 1.0 mL, 1 eq). The resulting mixture was stirred at
25.degree. C. for 17 hours to give a purple suspension. TLC
(PE/EtOAc=2/1) showed the reaction was completed. The mixture was
concentrated under reduced pressure to give 5.53 g (crude) of A39
as purple oil.
General Procedure for Synthesis A40
[0207] To a solution of A39 (5.53 g, 16.4 mmol, 1 eq) in MeOH (50
mL), was added polyformaldehyde (2.46 g, 16.39 mmol), HOAc (984 mg,
16.4 mmol, 937 uL, 1 eq) and NaBH.sub.3CN (1.03 g, 16.4 mmol, 1
eq). The resulting mixture was stirred at 25.degree. C. for 72
hours to give a light red solution. TLC (PE/EtOAc=1/1) showed the
reaction was completed. The reaction mixture was concentrated under
reduced pressure to give a residue. The residue was dissolved in
EtOAc (50 mL). Sat. Na.sub.2CO.sub.3 (40 mL) was added to the
solution. The aqueous was extracted with EtOAc (20 mL). The
combined organic layer were washed with brine (15 mL.times.2),
dried over anhydrous Na.sub.2SO.sub.4, filtered and concentrated
under reduced pressure to give 5.38 g (crude) of A40 as brown
oil.
General Procedure for Synthesis A41
[0208] To a solution of A40 (5.38 g, 15.3 mmol, 1 eq) in DCM (90
mL) was added TFA (46.2 g, 405 mmol, 30 mL, 26.5 eq). The resulting
mixture was stirred at 25.degree. C. for 16 hours to give a brown
mixture. TLC (PE/EtOAc=1/1) showed a little starting material was
still remained. LCMS (Rt=0.665 min) showed 83% of desired compound
MS. The reaction mixture was concentrated under reduced pressure.
The residue was dissolved in EtOAc (50 mL). Sat. Na.sub.2CO.sub.3
(100 mL) was added to the solution. The aqueous phase was extracted
with EtOAc (30 mL). The combined organic layer extract was washed
with brine (20 mL.times.3), dried over anhydrous Na.sub.2SO.sub.4,
filtered and concentrated under reduced pressure to give a residue.
The residue was purified by Combi flash (PE/EtOAc=1/1) to give 1.98
g (yield: 48.4%) of A41 as a brown gum.
General Procedure for Synthesis A42
[0209] To a solution of A41 (520 mg, 1.95 mmol, 1 eq) and D1-1 (827
mg, 4.86 mmol, 475 uL, 2.5 eq) in MeCN (10 mL) was added
K.sub.2CO.sub.3 (672 mg, 4.86 mmol, 2.5 eq) at 25.degree. C., then
the mixture was stirred at 60.degree. C. for 16 hours to give an
off-white suspension. TLC (PE/EtOAc=1/1) showed two new spots. LCMS
(Rt=0.711 min) showed the reaction was completed. The mixture was
concentrated in vacuum, and the residue was quenched by water (30
mL). The resulting solution was extracted with EtOAc (20
mL.times.3). The combined organic layers were washed with brine (10
mL.times.2), dried over anhydrous Na.sub.2SO.sub.4, filtered and
concentrated in vacuum to give a residue. The residue was purified
by Combi flash (PE/EtOAc=3/1 to 2/1 to 1/1) to give 280 mg (yield:
46.5%) of A42 as a yellow oil.
General Procedure for Synthesis A43
[0210] To a solution of A42 (280 mg, 0.905 mmol, 1 eq) in MeOH (4
mL), was added NaOH (3 M, 603 uL, 2 eq) in H.sub.2O (0.6 mL), and
the mixture was stirred at 20.degree. C. for 18 hours to give a
colorless mixture. TLC (PE/EtOAc=1/1) showed a new spot. LCMS
(Rt=0.518 min) showed the reaction was completed. The reaction
mixture was concentrated under reduced pressure to give a residue.
The residue was quenched by addition H.sub.2O (10 mL), and then
extracted with EtOAc (20 mL). Then 2 N hydrochloric acid was added
to that aqueous phase to adjust pH=1 by aq. HCl. The aqueous phase
was extracted with DCM (10 mL.times.8). The combined organic layers
were washed with brine (10 mL.times.3), dried over anhydrous
Na.sub.2SO.sub.4, filtered and concentrated in vacuum to give 90 mg
(crude) of A43 as a light yellow solid. LCMS showed 81% purity. The
aqueous phase still contained product from LCMS. Then toluene (10
mL.times.5) was added to the aqueous phase. The aqueous phase was
concentrated under reduced pressure to give 130 mg (crude, contain
NaCl salt) of A43 as a white solid. LCMS showed 98% of desired MS
value.
General Procedure for Synthesis 34
[0211] To a solution of A43 (130 mg, 0.457 mmol, 3 eq) and B1-1 (45
mg, 0.15 mmol, 1 eq) in pyridine (1.5 mL), was added EDCI (44 mg,
0.23 mmol, 1.5 eq). The resulting mixture was stirred at 20.degree.
C. for 16 hours to give a yellow mixture. TLC (EtOAc) showed a new
spot. LCMS showed 4.9% of desired compound MS. The mixture was
stirred at 20.degree. C. for another 8 hours. Then additional EDCI
(44 mg, 0.23 mmol, 1.5 eq) was added. The mixture was stirred at
20.degree. C. for 48 hours. LCMS (Rt=0.773 min) showed 7.2% of
desired compound MS. The mixture was partitioned between DCM (50
mL) and water (30 mL). The aqueous phase was extracted with DCM (20
mL.times.3). The combined extracts were washed with saturated brine
(10 mL.times.2), dried over anhydrous Na.sub.2SO.sub.4, filtered
and concentrated under reduced pressure to give a residue. The
residue was purified by prep-TLC
[(PE/EtOAc=1/1)/(DCM/MeOH=10/1)=1/1] to give 34 (3.7 mg, yield:
4.2%, purity: 97.5%) as a white powder.
##STR00198##
General Procedure for Synthesis B3
[0212] A suspension of Na (2.66 g, 116 mmol, 3 eq) in MeOH (40 mL)
was stirred at 25.degree. C. for 30 min. Then B2 (10 g, 38.6 mmol,
1 eq) was added into the above mixture. The reaction was stirred at
80.degree. C. for 17 hours to give a yellow suspension. TLC showed
starting material was not consumed completely. The mixture was
cooled to room temperature. The mixture was adjusted pH to 1-2 by
aqueous HCl (4 M), extracted with EtOAc (200 mL.times.2). The
combined extract was washed with brine (200 mL.times.2), dried over
anhydrous Na.sub.2SO.sub.4 and filtered, then concentrated under
reduced pressure to give B3 (8.3 g, yield: 79%) as yellow oil.
General Procedure for Synthesis B4
[0213] To a solution of B3 (7.1 g, 26.2 mmol, 1 eq) and
diphenylmethanimine (5.7 g, 31.4 mmol, 1.2 eq) in toluene (150 mL)
was added Pd.sub.2(dba).sub.3 (599 mg, 0.655 mmol, 0.025 eq), BINAP
(1.22 g, 1.96 mmol, 0.075 eq) and t-BuONa (3.52 g, 36.7 mmol, 1.4
eq) under N.sub.2. The resulting mixture was heated at 90.degree.
C. and stirred for 12 hours to give red solution. TLC showed
desired product was formed. The reaction mixture was quenched by
addition H.sub.2O (150 mL) and extracted with EtOAc (150
mL.times.2). The combined organic layers were washed with brine (50
mL.times.2), dried over Na.sub.2SO.sub.4, filtered and concentrated
under reduced pressure to give a residue. The residue was purified
by column chromatography (SiO.sub.2, PE/EtOAc=20:1) to obtain 6 g
crude product, which was triturated with PE/EtOAc=40 mL/2 mL to
give 4.65 g of B4 as a light yellow solid.
General Procedure for Synthesis B5
[0214] To a solution of B4 (5.4 g, 14.5 mmol, 1 eq) in HO/MeOH (4M)
(20 mL) was stirred at 20.degree. C. for 30 min to give white
suspension. LCMS showed the reaction was completed. The reaction
mixture was concentrated under reduced pressure to give a residue.
Then diluted with H.sub.2O (50 mL) and basified with sat.
NaHCO.sub.3 solution to pH=.about.9, extracted with EtOAc (50 mL).
The combined organic layers were washed with brine (10 mL), dried
over Na.sub.2SO.sub.4, filtered and concentrated under reduced
pressure to give a residue. The residue was purified by column
chromatography (SiO.sub.2, PE/EtOAc=3/1) to give B5 (2.5 g, yield:
83%) as a yellow oil.
General Procedure for Synthesis B6
[0215] A mixture of 2,2-dimethyl-1,3-dioxane-4,6-dione (1.83 g,
12.7 mmol, 1.05 eq) and diethoxymethoxyethane (1.97 g, 13.3 mmol,
1.1 eq) was heated at 50.degree. C. and stirred for 1.5 hours. Then
a solution of B5 (2.5 g, 12.1 mmol, 1 eq) in MeOH (10 mL) was added
to the mixture. The mixture was stirred at 50.degree. C. for 1 hour
to give yellow suspension. The mixture was cooled to room
temperature and added EtOH (20 mL). The resulting mixture was
stirred at 0.degree. C. for 30 min. White powder was observed. The
mixture was filtered. The filter cake was washed with EtOH (20 mL)
and dried over high vacuum to give 1.3 g as a white solid, However
LCMS and H NMR showed it is not the desired product. The filtrate
was extracted with EtOAc (50 mL). The combined organic layers were
washed with brine (20 mL), dried over Na.sub.2SO.sub.4, filtered
and concentrated under reduced pressure to give a residue. The
residue was purified by silica column (PE/EtOAc=3/1) to obtain B6
(550 mg, crude) as a yellow solid. H NMR showed the mixture of
desired product and B5.
General Procedure for Synthesis B7
[0216] To a solution of B6 (550 mg, 1.00 mmol, 1 eq) in Dowtherm A
(4 mL) was heated at 210.degree. C. and stirred for 1 hour under
N.sub.2 to give brown solution. TLC showed the reaction was
completed. The reaction was cooled to room temperature and quenched
by addition H.sub.2O (20 mL) and extracted with EtOAc (20
mL.times.2). The combined organic layers were washed with brine (10
mL.times.2), dried over Na.sub.2SO.sub.4, filtered and concentrated
under reduced pressure to give a residue. The residue was purified
by column chromatography (SiO2, DCM:MeOH=10:1) to give B7 (30 mg,
yield: 11%, purity: 98%) as a yellow solid.
General Procedure for Synthesis B8
[0217] To a solution of B7 (30 mg, 0.116 mmol, 1 eq) and
1,2-difluoro-4-nitro-benzene (18.4 mg, 0.116 mmol, 1 eq) in DMF (1
mL) was added Cs.sub.2CO.sub.3 (56.6 mg, 0.174 mmol, 1.5 eq). The
resulting mixture was heated at 40.degree. C. and stirred for 2 hrs
to give red solution. LCMS showed the reaction was completed. The
reaction was quenched by addition H.sub.2O (20 mL) and extracted
with EtOAc (20 mL.times.2). The combined organic layers were washed
with brine (10 mL.times.2), dried over Na.sub.2SO.sub.4, filtered
and concentrated under reduced pressure to give a residue. The
residue was purified by prep-TLC (PE:EtOAc=1:1) to give B8 (30 mg,
yield: 65%) as a yellow solid.
General Procedure for Synthesis B1-2
[0218] To a solution of B8 (30 mg, 0.075 mmol, 1 eq) in EtOH (5 mL)
was added Pd/C (wet) in 50% water (20 mg, 10% purity) under
N.sub.2. The suspension was degassed under vacuum and purged with
H.sub.2 several times. The mixture was stirred under H.sub.2 (15
psi) at 20.degree. C. for 12 hours to give black suspension. LCMS
and TLC showed part of the starting material remain and one new
spot was formed. The reaction mixture was filtered and the filter
was concentrated. The residue was purified by prep-TLC
(PE:EtOAc=1:1) to give B1-2 (20 mg, yield: 72%, purity: 100%) as an
off-white solid.
General Procedure for Synthesis A44
[0219] To a suspension of A18 (1.43 g, 7.21 mmol, 1 eq),
K.sub.2CO.sub.3 (2.49 g, 18 mmol, 2.5 eq) in MeCN (143 mL) was
added D1-1 (3.07 g, 18 mmol, 2.5 eq) at 25.degree. C. The reaction
was stirred at 60.degree. C. for 17 hours to give a yellow
suspension. LCMS showed the reaction was completed. The reaction
mixture was worked up after combining with the pilot reaction from
100 mg of A18. The reaction mixture was partitioned between EtOAc
(200 mL) and H.sub.2O (200 mL). The organic layer was washed with
brine (200 mL.times.2), dried over anhydrous Na.sub.2SO.sub.4,
filtered and concentrated under reduced pressure to give black oil.
The crude product was purified by combi flash (EtOAc:PE=0:1 to 1:4)
to give A44 (0.8 g, yield: 43%, purity: 95%) as yellow oil.
General Procedure for Synthesis A45
[0220] To a solution of A44 (0.8 g, 3.33 mmol, 1 eq) in MeOH (10
mL) was added NaOH (400 mg, 9.99 mmol, 3 eq) in H.sub.2O (3 mL).
The reaction was stirred at 25.degree. C. for 1.5 hours to give a
yellow solution. LCMS showed the starting material was not consumed
completely. The reaction was stirred at 25.degree. C. for 17 hours
to give a yellow solution. LCMS showed the starting material was
consumed completely. The reaction was adjusted to pH=4, and
extracted with EtOAc (200 mL). The organic layer was washed with
brine (150 mL), dried over anhydrous Na.sub.2SO.sub.4, filtered and
concentrated under reduced pressure to give A45 (710 mg, crude) as
yellow oil.
General Procedure for Synthesis 35
[0221] To a solution of B1-2 (20 mg, 1 eq) and A45 (17.3 mg, 0.081
mmol, 1.5 eq) in pyridine (0.5 mL) was added EDCI (15.6 mg, 0.081
mmol, 1.5 eq). The resulting mixture was stirred at 20.degree. C.
for 12 hrs to give yellow solution. LCMS showed the reaction was
completed. The reaction mixture concentrated under reduced pressure
to give a residue. The residue was purified by prep-HPLC (column:
Waters Xbridge 150*50 10u; mobile phase: [water (0.05% ammonia
hydroxide v/v)-ACN]; B %: 55%-85%, 7.8 min), then concentrated and
lyophilized to give 35 (11.8 mg, yield: 38%, purity: 100%) as a
white powder.
##STR00199##
General Procedure for Synthesis A46
[0222] A mixture of D1-6 (831.50 mg, 5.04 mmol, 1.20 eq), A6 (1.00
g, 4.20 mmol, 1.00 eq) and Cs.sub.2CO.sub.3 (2.74 g, 8.40 mmol,
2.00 eq) in dioxane (8.5 mL) was heated to 80.degree. C. for 12 h.
TLC (petroleum ether/ethyl acetate=1/1, A6: Rf=0.42, A46: Rf=0.36)
showed A6 was consumed. The mixture was concentrated and the
filtrate was concentrated. The residue was purified by column
chromatography (SiO.sub.2, petroleum ether/ethyl acetate=3/1 to
1/1) (TLC: petroleum ether/ethyl acetate=1/1, A46: Rf=0.36). A46
(0.82 g, 2.54 mmol, 60.60% yield) was obtained as yellow oil.
General Procedure for Synthesis A47
[0223] To a solution of A46 (0.82 g, 2.54 mmol, 1.00 eq) in MeOH
(5.7 mL) was added dropwise a solution of NaOH (305 mg, 7.63 mmol,
3.00 eq) in H.sub.2O (1.64 mL) at 15.about.25.degree. C. The
mixture was stirred at 15.about.25.degree. C. for 5 h. TLC
(petroleum ether/ethyl acetate=1/1, A46: Rf=0.34, A47: Rf=0.01)
showed A46 was consumed completely. The mixture was concentrated.
The pH value of the residue was adjusted to 5.about.6 with 4 M HCl
(15.0 mL). The mixture was extracted with a solution of DCM and IPA
(3/1, 20 mL.times.4). The combined organic layers were dried over
anhydrous Na.sub.2SO.sub.4, filtered and concentrated. A47 (0.68 g,
2.31 mmol, 90.83% yield) was obtained as yellow oil.
General Procedure for Synthesis 44
[0224] To a solution of A47 (0.15 g, 510 umol, 1.00 eq) in pyridine
(5 mL) was added EDCI (147 mg, 765 umol, 1.50 eq) stir for 15 min.
The B1-1 (155 mg, 520 umol, 1.02 eq) was added the mixture. The
mixture was stirred at 25.degree. C. for 12 h. LC-MS (44: RT=0.84
min) showed A47 was consumed completely and one main peak with
desired mass was detected. HPLC (ET24988-44-P1A, compound 44:
RT=2.55 min) showed one main peak was detected. The mixture was
concentrated under reduced pressure to give a residue. The residue
was purified by reversed-phase HPLC (0.1% HCl condition). 44 (0.08
g, 134 umol, 26.3% yield, 96.2% purity) was obtained as a yellow
solid.
##STR00200##
General Procedure for Synthesis D1-8
[0225] To a suspension of Mg (67.1 g, 2.76 mol, 10 eq) and I.sub.2
(701 mg, 2.76 mmol, 0.01 eq) in THF (350 mL) was added slowly
1,2-dibromoethane (259 g, 1.38 mol, 5.0 eq) in THF (1 L) slowly at
a rate as to keep the internal temperature between 40-55.degree. C.
After the addition, a solution of D1-7 (50 g, 276 mmol, 1 eq) in
THF (375 mL) was added drop-wise. The reaction mixture was kept at
40-55.degree. C. for 16 h. TLC (petroleum ether/ethyl acetate=2/1,
D1-8: Rf=0.27) indicated D1-7 was consumed completely and many new
spots formed. The reaction was quenched with saturated NH.sub.4Cl
(3 L) slowly at 0.degree. C. The mixture was extracted and the
aqueous phase was extracted with DCM/i-PrOH (v/v=10/1, 2.5 L and 1
L). The combined organic layers were dried over Na.sub.2SO.sub.4,
filtered and concentrated. The residue was purified by column
chromatography (SiO.sub.2, petroleum ether/ethyl acetate=10/1 to
3/1). D1-8 (14 g, 137 mmol, 49.6% yield) was obtained as yellow
oil, which was used into next step without further
purification.
General Procedure for Synthesis D1-9
[0226] To the solution of D1-8 (7 g, 68.5 mmol, 1.0 eq) and TEA
(20.8 g, 206 mmol, 3.0 eq) in DCM (60 mL) was added MsCl (15.7 g,
137.1 mmol, 2.0 eq) slowly drop-wise at 0.degree. C. for 0.5 h. The
mixture was stirred at 25.degree. C. for 3 h. TLC (petroleum
ether/ethyl acetate=2/1, PMA, D1-8: Rf=0.27) showed D1-8 was
consumed completely. The reaction mixture was quenched by addition
of aqueous NH.sub.4Cl (5 mL), extracted with CH.sub.2Cl.sub.2 (30
mL, 15 mL). The organic phases were combined and dried over
Na.sub.2SO.sub.4, filtered and concentrated. The residue was
purified by column chromatography (SiO.sub.2, petroleum ether/ethyl
acetate=20/1 to 1/1). D1-9 (11.5 g, crude) was obtained as red oil,
which was used into next step without further purification.
General Procedure for Synthesis A48
[0227] To the solution of D1-9 (21.3 g, 89.43 mmol, 1.0 eq) and
Cs.sub.2CO.sub.3 (87.42 g, 268.30 mmol, 3.0 eq) in dioxane (150 mL)
was added A6 (20.95 g, 116.26 mmol, 1.3 eq) at 15.degree. C. The
mixture was stirred at 40.degree. C. for 3 h. TLC (petroleum
ether/ethyl acetate=1/1, A47: Rf=0.3) showed D1-9 was consumed
completely. The reaction mixture was added water (5 mL) and
extracted with EtOAc (5 mL). The residue was purified by column
chromatography (SiO.sub.2, petroleum ether/ethyl acetate=20/1 to
3/1). A48 (15.2 g, crude) was obtained as red oil, which was used
into the next step without further purification.
General Procedure for Synthesis A49
[0228] To the solution of A48 (15.5 g, 48.1 mmol, 1.0 eq) in MeOH
(30 mL) and H.sub.2O (30 mL) was added LiOH.H.sub.2O (6.05 g, 144
mmol, 3 eq) at 15.degree. C. The mixture was stirred at 15.degree.
C. for 3 h. TLC (petroleum ether/ethyl acetate=1/1, A47: Rf=0.3)
showed A47 was consumed completely. The reaction mixture was
concentrated to remove MeOH. The pH value was adjusted to 2-3 with
HCl solution (1M, 18 mL). The mixture was extracted with DCM (100
mL). The organic phase was dried over Na.sub.2SO.sub.4. The mixture
was filtered and the filtrate was concentrated to get product. A49
(14 g, 47.6 mmol, 98.9% yield) was obtained as a light red solid,
which was used into next step without further purification.
General Procedure for Synthesis 120
[0229] To the solution of A49 (200 mg, 679 umol, 1 eq) in DCM (2
mL) was added B1-3 (243 mg, 815 umol, 1.2 eq) and DIPEA (175 mg,
1.36 mmol, 2.0 eq) at 25.degree. C. To the mixture was added T3P
(648 mg, 1.02 mmol, 50% purity in EtOAc solution, 1.5 eq) at
15.degree. C. The mixture was stirred at 25.degree. C. for 16 h.
LCMS (120: RT=1.01 min) showed that 12.9% of 120 was detected. The
mixture was washed with water (2 mL). The organic phase was
concentrated to get a residue. The residue was purified by
prep-HPLC (column: Waters Xbridge Prep OBD C18 150*40 mm*10 um;
mobile phase: [water (10 mM NH4HCO3)-ACN]; B %: 30%-50%, 8 min).
120 (40 mg, 68.7 umol, 10.1% yield, 98.6% purity) was obtained as
an off-white solid.
##STR00201##
General Procedure for Synthesis B10
[0230] A mixture of B9 (500 mg, 2.24 mmol, 1 eq) and
3-fluoro-4-nitro-phenol (703 mg, 4.47 mmol, 2 eq) in chlorobenzene
(10 mL) was stirred at 131.degree. C. for 12 hours to give an
off-white suspension. LCMS (Rt=1.045-1.107 min) showed .about.25%
of 545-1 and .about.75% of desired MS value. The reaction mixture
were filtered, filtrate cake was washed with toluene (10 mL) and
dried under vacuum to give a crude compound. The crude compound was
basified with 10% aq.NaOH, and the suspension was stirred for 1
hour at room temperature. The mixture was filtered, filtrate cake
was dried in high vacuum to give B10 (300 mg, 37.6% yield, 96.5%
purity) as a yellow solid.
General Procedure for Synthesis B1-4
[0231] To a mixture of B10 (300 mg, 0.871 mmol, 1 eq) and
NH.sub.4Cl (466 mg, 8.71 mmol, 10 eq) in EtOH (10 mL) was added Zn
(570 mg, 8.71 mmol, 10 eq). The reaction mixture was stirred at
20.degree. C. for 12 hours to give a black mixture. LCMS (Rt=0.997
min) showed the reaction was completed. The mixture was filtered
and the filtrate was concentrated under reduced pressure to give
B1-4 (270 mg, 99% yield) as a yellow solid. The crude product was
used for the next step without further purification.
General Procedure for Synthesis 115
[0232] To a solution of compound A49 (1.00 eq) and B1-4 (1.02 eq)
in pyridine (5 V) was added EDCI (1.50 eq) at 15.about.25.degree.
C. under N.sub.2. The mixture was stirred for at
15.about.25.degree. C. 12 h under N.sub.2. LCMS showed A49 was
consumed completely and one main peak with desired m/z was
detected. The mixture was concentrated under reduced pressure to
give a residue. The crude product was purified by reversed-phase
HPLC. 115 (128 mg, 60.7% yield, 95.6% purity) was obtained as a
white solid.
##STR00202##
General Procedure for Synthesis A50
[0233] A mixture of compound A6 (3.00 g, 12.6 mmol, 1.00 eq), D1-10
(2.78 g, 15.1 mmol, 1.74 mL, 1.20 eq) and Cs.sub.2CO.sub.3 (8.21 g,
25.2 mmol, 2.00 eq) in dioxane (45 mL) was heated to 30.degree. C.
for 60 h. LCMS (A49: RT=1.16 min) showed A6 was consumed completely
and the desired MS was detected. The mixture was filtered and the
filtrate was concentrated. The residue was purified by column
chromatography (SiO.sub.2, petroleum ether/ethyl acetate=3/1 to
1/1) (TLC: petroleum ether/ethyl acetate=1/1, A49: Rf=0.50). A50
(2.5 g, 8.50 mmol, 67.4% yield) was obtained as yellow solid.
General Procedure for Synthesis A51
[0234] To a solution of A49 (2.5 g, 8.50 mmol, 1 eq) in MeOH (17.5
mL) and H.sub.2O (5 mL) was added NaOH (1.02 g, 25.5 mmol, 3 eq).
The mixture was stirred at 25.degree. C. for 4 hr. TLC (petroleum
ether/ethyl acetate=3/1, A49: Rf=0.30, A50: Rf=0.00) indicated A49
was consumed completely and one major new spot with larger polarity
was detected. The reaction mixture was added ethyl acetate (20 mL)
and H.sub.2O (10 mL) to stirred at 15.about.25.degree. C. for 10
min. The mixture was separated. The pH value of the aqueous layer
was adjusted to 5.about.6 with 4M HCl (10.0 mL). The mixture was
extracted with a solution of DCM and IPA (3/1, 30 mL.times.4). The
combined organic layers were dried over anhydrous Na.sub.2SO.sub.4,
filtered and concentrated. A51 (2 g, 7.51 mmol, 88.4% yield) was
obtained as a light yellow oil
General Procedure for Synthesis B12
[0235] To a mixture of B11 (1 g, 4.87 mmol, 1 eq) and
1,2-difluoro-4-nitro-benzene (1.55 g, 9.75 mmol, 1.08 mL, 2 eq) in
DMSO (10 mL) was added Cs.sub.2CO.sub.3 (3.18 g, 9.75 mmol, 2 eq).
The reaction mixture was stirred at 50.degree. C. for 12 hours to
give a brown mixture. LCMS showed the reaction was completed. The
mixture was allowed to 20.degree. C., diluted with EtOAc (100 mL),
washed with water (50 mL.times.3) and brine (80 mL), dried over
anhydrous Na.sub.2SO.sub.4, filtered and concentrated to give a
crude compound. The crude compound was purified by flash silica gel
chromatography (ISCO.RTM.; 20 g SepaFlash.RTM. Silica Flash Column,
Eluent of 10%.about.50%.about.100% Ethyl acetate/Petroleum ether
gradient @ 30 mL/min) to afford B12 (1 g, 59% yield, 98.9% purity)
as a yellow solid.
General Procedure for Synthesis B1-5
[0236] To a mixture of B12 (1 g, 2.90 mmol, 1 eq) in MeOH (20 mL)
was added Pd/C (200 mg, 10% purity, 50% H.sub.2O) under N.sub.2.
The suspension was degassed under vacuum and purged with H2 twice.
The mixture was stirred under H.sub.2 (15 psi) at 20.degree. C. for
12 hours to give a black mixture. LCMS showed the reaction was
completed. The mixture was filtered and the filtrate was
concentrated under reduced pressure to give B1-5 (560 mg, 61.3%
yield) as an off-white solid. The product was used for the next
step without further purification.
General Procedure for Synthesis 53
[0237] To a solution of A50 (1.00 eq) and B1-5 (1.02 eq) in
pyridine (7 V) was added EDCI (1.50 eq) at 15.about.25.degree. C.
under N.sub.2. The mixture was stirred for at 15.about.25.degree.
C. 12 h under N.sub.2. LCMS showed A51 was consumed completely and
one main peak with desired m/z was detected. The mixture was
concentrated under reduced pressure to give a residue. The crude
product was purified by reversed-phase HPLC. 53 (199 mg, 61.7%
yield, 98.3% purity) was obtained as a yellow solid.
##STR00203##
General Procedure for Synthesis B13
[0238] To a mixture of B11 (500 mg, 2.44 mmol, 1 eq) and
1-fluoro-2-methyl-4-nitro-benzene (756 mg, 4.87 mmol, 2 eq) in DMSO
(5 mL) was added Cs.sub.2CO.sub.3 (1.59 g, 4.87 mmol, 2 eq). The
reaction mixture was stirred at 50.degree. C. for 12 hours to give
a black mixture. LCMS showed the reaction was completed. The
mixture was diluted with EtOAc (100 mL), washed with water (30
mL.times.3) and brine (30 mL), concentrated under reduced pressure
to give a crude compound. The crude compound was purified by flash
silica gel chromatography (ISCO.RTM.; 12 g SepaFlash.RTM. Silica
Flash Column, Eluent of 5-70% Ethyl acetate/Petroleum ether
gradient @ 30 mL/min) to afford B13 (190 mg, 22.4% yield, 98%
purity) as a yellow solid.
General Procedure for Synthesis B1-6
[0239] To a mixture of B13 (190 mg, 558.28 umol, 1 eq) in MeOH (10
mL) was added Pd/C (38 mg, 10% purity, 50% H.sub.2O) under N.sub.2.
The suspension was degassed under vacuum and purged with H2 two
times. The mixture was stirred under H.sub.2 (15 psi) at 20.degree.
C. for 12 hours to give a black mixture. LCMS (Rt=0.670 min) showed
the reaction was completed. The reaction mixture was filtered and
filtrate was concentrated under reduced pressure to give B1-6 (150
mg, crude) as a yellow oil. The crude compound was used for the
next step without further purification.
General Procedure for Synthesis 58
[0240] To a solution of A51 (1.00 eq) and B1-6 (1.02 eq) in
pyridine (7 V) was added EDCI (1.50 eq) at 15.about.25.degree. C.
under N.sub.2. The mixture was stirred for at 15.about.25.degree.
C. 12 h under N.sub.2. LCMS showed A51 was consumed completely and
one main peak with desired m/z was detected. The mixture was
concentrated under reduced pressure to give a residue. The crude
product was purified by reversed-phase HPLC. 58 (168 mg, 52.4%
yield, 98.2% purity) was obtained as a white solid.
##STR00204##
General Procedure for Synthesis A53
[0241] To EtOH (25 mL) was added Na (339.38 mg, 14.76 mmol, 349.88
uL, 1.1 eq), after Na disappeared, diethyl propanedioate (3.01 g,
18.79 mmol, 2.84 mL, 1.4 eq) was added into the above mixture, and
the mixture was stirred at 100.degree. C. for 30 min. To the
reaction mixture was added a solution of A52 (2.00 g, 13.42 mmol,
1.50 mL, 1 eq) in EtOH (10 mL) dropwised over 10 min, and then the
mixture was stirred for 17 hr. TLC indicated that one major new
spot with lower polarity was detected. The reaction mixture was
concentrated under reduced pressure to remove the solvent. The
residue was diluted with water (50 mL) and extracted with EtOAc (50
mL*2). The combined organic layers were washed with HCl (1N) (25
mL*2), dried over Na.sub.2SO.sub.4, filtered and concentrated under
reduced pressure to give a residue. The residue was purified by
flash silica gel chromatography (Eluent of 0-2% Ethyl
acetate/Petroleum ether gradient) to give 2.6 g (yield: 84.87%) of
A53 as colorless oil.
General Procedure for Synthesis A54
[0242] To the solution of A53 (2.6 g, 11.39 mmol, 1 eq) in H.sub.2O
(10 mL) and EtOH (10 mL) was added KOH (2.56 g, 45.56 mmol, 4 eq),
and the mixture was stirred at 15.degree. C. for 1.5 hr. TLC
indicated the reaction was completed. The reaction mixture was
concentrated under reduced pressure to remove the solvent. The
residue was diluted with water (20 mL) and adjust pH to 3-5 by HCl
(1N), and then it was extracted with EtOAc (20 mL*3). The combined
organic layers were washed with aqueous NaCl (20 mL*3), dried over
Na.sub.2SO.sub.4, filtered and concentrated under reduced pressure
to give 1.7 g (yield: 86.69%) of A54 as a white solid.
General Procedure for Synthesis A55
[0243] A solution of A54 (1.7 g, 9.87 mmol, 1 eq) in NMP (2 mL) was
stirred at 110.degree. C. for 2 hr. TLC indicated that material was
remained, and one major new spot with lower polarity was detected.
Then the mixture was heated to 120.degree. C. for 17 hr, and TLC
indicated that material was still remained. The reaction mixture
was diluted with water (30 mL) and it was extracted with EtOAc (30
mL*2). The combined organic layers were washed with aqueous NaCl
(30 mL*2), dried over Na.sub.2SO.sub.4, filtered and concentrated
under reduced pressure to give 1.01 g (yield: 79.81%) of A55 as a
white solid.
General Procedure for Synthesis A56
[0244] To the solution of A55 (1.01 g, 7.88 mmol, 1 eq) and
2,2-dimethyl-1,3-diozane-4,6-dione (1.25 g, 8.67 mmol, 1.1 eq) in
DCM (15 mL) was added DMAP (1.44 g, 11.82 mmol, 1.5 eq), after
cooling the mixture to 0.degree. C., EDCI (2.11 g, 11.03 mmol, 1.4
eq) was added. The mixture was stirred at 15.degree. C. for 17 hr.
TLC indicated that the reaction was completed and one major new
spot with lower polarity was detected. The mixture was treated with
50 ml of water and the mixture was extracted with EtOAc (50
mL.times.2). The combined organic layer was washed with 1N HCl (20
mL.times.2), dried over magnesium sulfate, and concentrated under
reduced pressure to give 1.9 g (yield: 94.82%) of A56 as yellow
oil.
General Procedure for Synthesis A57
[0245] A solution of A56 (1.9 g, 7.47 mmol, 1 eq) in EtOH (40 mL)
was stirred at 80.degree. C. for 4 hr. TLC indicated that the
reaction was completed. The reaction mixture concentrated under
reduced pressure to give 1.47 g (yield: 99.23%) of A57 as yellow
oil.
General Procedure for Synthesis A58
[0246] A solution of NaNO.sub.2 (767.36 mg, 11.12 mmol, 1.5 eq) in
H.sub.2O (12 mL) was added dropwise to the solution of A57 (1.47 g,
7.41 mmol, 1 eq) in CH.sub.3COOH (4 mL) and H.sub.2O (12 mL) under
0.degree. C. The stirring mixture was maintained at 0.degree. C.
for 2 h and then at 15.degree. C. for 2.5 h. Then, water (10 ml)
was added and the mixture was stirred for 17 hr. TLC indicated the
reaction was completed. The reaction mixture was diluted with water
(10 mL) and it was extracted with EtOAc (30 mL*2). The combined
organic layers were washed with sat. aq. NaHCO.sub.3 (15 mL*2),
brine (15 mL*2), dried over Na.sub.2SO.sub.4, filtered and
concentrated under reduced pressure to give 1.42 g (yield: 84.27%)
of A58 as yellow oil.
General Procedure for Synthesis A59
[0247] To the solution of A58 (3.1 g, 13.64 mmol, 1 eq) in EtOH (50
mL) were added wet Pd/C (0.4 g) and conc. HCl (227.82 mg, 6.25
mmol, 223.36 uL). The suspension was degassed and purged with
H.sub.2 for 3 times. The mixture was stirred under H.sub.2 (15 Psi)
at 40.degree. C. for 17 hr. TLC indicated the reaction was
completed. The suspension was filtered through a pad of Celite and
the pad was washed with EtOH (10 mL.times.3). The combined
filtrates were concentrated to dryness to 3.3 g (yield: 96.87%,
HCl) of A59 as a white solid.
General Procedure for Synthesis A60
[0248] To the solution of A59 (1.00 g, 4.00 mmol, 1 eq, HCl) in DCM
(5 mL) was added TEA (1.22 g, 12.01 mmol, 1.67 mL, 3 eq), and then
4,4,4-trifluorobutanoyl chloride (1.29 g, 8.01 mmol, 2 eq) was
added dropwise at 0.degree. C. After addition, the mixture was
stirred at 20.degree. C. for 1 hr to give a yellow mixture. TLC
indicated the reaction was completed. The reaction mixture was
diluted with water (50 mL) and it was extracted with EtOAc (50
mL*2). The combined organic layers were washed with sat. aq.
NaHCO.sub.3 (20 mL*2), NaCl (20 mL*2), dried over Na.sub.2SO.sub.4,
filtered and concentrated under reduced pressure to give a residue.
The residue was purified by flash silica gel chromatography (Eluent
of 0-20% Ethyl acetate/Petroleum ether) to afford 0.32 g (yield:
23.69%) of A60 as yellow oil.
General Procedure for Synthesis A61
[0249] To the solution of A60 (0.32 g, 948.62 umol, 1 eq) in DCM
(10 mL) were added PPh.sub.3 (497.62 mg, 1.90 mmol, 2 eq), I.sub.2
(481.53 mg, 1.90 mmol, 382.17 uL, 2 eq) and TEA (383.96 mg, 3.79
mmol, 528.15 uL, 4 eq). The mixture was stirred at 20.degree. C.
for 2 hr. TLC indicated the reaction was completed. The reaction
mixture was diluted with water (20 mL) and it was extracted with
EtOAc (10 mL*3). The combined organic layers were washed with
aqueous NaCl (10 mL*3), dried over Na.sub.2SO.sub.4, filtered and
concentrated under reduced pressure to give a residue. The residue
was purified by flash silica gel chromatography (Silica Flash
Column, Eluent of 0-20% Ethyl acetate/Petroleum ether gradient) to
afford 0.22 g (yield: 72.63%) of A61 as yellow oil.
General Procedure for Synthesis A62
[0250] To a solution of compound A61 (0.29 g, 908.18 umol, 1 eq) in
THF (8 mL) and H.sub.2O (2 mL) was added LiOH.H.sub.2O (57.17 mg,
1.36 mmol, 1.5 eq), and the mixture was stirred at 20.degree. C.
for 17 hr to give a yellow mixture. And then the mixture was
stirred at 50.degree. C. for another 6 hr. LCMS showed the reaction
was completed. The residue was diluted with water (20 mL) and then
lyophilized to afford 0.325 g (crude) of A62 as a white solid.
General Procedure for Synthesis 176
[0251] To the solution of A62 (0.15 g, crude) in pyridine (2 mL)
were added B1-1 (153.11 mg, 514.99 umol, 1 eq) and EDCI (148.09 mg,
772.49 umol, 1.5 eq). The mixture was stirred at 15.degree. C. for
17 hr to give a yellow mixture. LCMS showed 7% of desired product
was detected. The reaction mixture was diluted with water (30 mL)
and it was extracted with EtOAc (10 mL*3). The combined organic
layers were washed with aqueous NaCl (10 mL*3), dried over
Na.sub.2SO.sub.4, filtered and concentrated under reduced pressure
to give a residue. The residue was purified by prep-TLC
(DCM:MeOH=20:1) to afford 5 mg (yield: 1.46%) of 176 as a white
solid.
##STR00205##
General Procedure for Synthesis A64
[0252] To a mixture of A63 (2 g, 9.17 mmol, 1 eq) in DMF (20 mL)
were added Cs.sub.2CO.sub.3 (7.47 g, 22.9 mmol, 2.5 eq) and
2,2,2-trifluoroethyl trifluoromethanesulfonate (4.26 g, 18.3 mmol,
2 eq) at 0.degree. C., the mixture was stirred at 15.degree. C. for
1 hour to give a pale mixture. LCMS showed the desired product was
observed. The mixture was poured into ice-water (30 mL). The
aqueous phase was extracted with ethyl acetate (20 mL.times.3). The
combined organic phase was washed with brine (50 mL.times.5), dried
with anhydrous Na.sub.2SO.sub.4, filtered and concentrated in
vacuum to afford crude product. The crude product was purified by
combi flash (PE/EtOAc=I/O to 3/1) to afford A63 (1.49 g, 42.5%
yield) as a white solid.
General Procedure for Synthesis A65
[0253] To a mixture of A64 (1.49 g, 3.90 mmol, 1 eq) and D1-3 (1.00
g, 11.70 mmol, 3 eq) in H.sub.2O (2 mL) and toluene (20 mL) were
added K.sub.3PO.sub.4 (2.07 g, 9.75 mmol, 2.5 eq), PCy.sub.3 (109
mg, 390 umol, 126 uL, 0.1 eq) and Pd(OAc).sub.2 (87.5 mg, 390 umol,
0.1 eq), and the mixture was stirred at 80.degree. C. under N.sub.2
protection for 5 hours to give a yellow mixture. LCMS showed the
desired product was observed. The mixture was partitioned between
EtOAc (15 mL) and water (10 mL). The aqueous phase was extracted
with EtOAc (10 mL.times.2). The combined organic extract was washed
with brine (10 mL), dried over Na.sub.2SO.sub.4, filtered,
concentrated under reduced pressure to give crude product. The
crude product was purified by combi flash (PE/EtOAc=I/O to 5/1 to
3/1) to afford A65 (150 mg, 11.2% yield) as a yellow solid.
General Procedure for Synthesis A66
[0254] To a mixture of A65 (150 mg, 437 umol, 1 eq) in THF (2 mL)
and H.sub.2O (1 mL) was added LiOH.H.sub.2O (36.6 mg, 874 umol, 2
eq), and the mixture was stirred at 15.degree. C. for 1 hours to
give a yellow mixture. LCMS showed the reactant was consumed. The
mixture was acidified with 1N HCl to pH=4-5, and then it was
partitioned between DCM (15 mL) and water (10 mL). The aqueous
phase was extracted with DCM (10 mL.times.2). The combined organic
extract was washed with brine (10 mL), dried over Na.sub.2SO.sub.4,
filtered, concentrated under reduced pressure to afford A66 (70 mg,
61.3% yield) as a yellow gum.
General Procedure for Synthesis 178
[0255] To a mixture of A66 (70 mg, 268 umol, 1 eq) and B1-1 (71.7
mg, 241 umol, 0.9 eq) in pyridine (2 mL) was added EDCI (102 mg,
535 umol, 2 eq), and the mixture was stirred at 15.degree. C. for
16 hours to give a yellow mixture. LCMS showed the reaction was
completed. The mixture was concentrated under reduced pressure to
give a crude product. The crude product was purified by prep-HPLC
(Waters Xbridge BEH C18 100*25 mm*5 um; mobile phase: [water (0.05%
ammonia hydroxide v/v)-ACN]; B %: 45%-75%, 9.5 min) to afford 178
(35.1 mg, 24.2% yield) as a white powder.
##STR00206##
General Procedure for Synthesis A68
[0256] To a suspension of A67 (4 g, 25.3 mmol, 1 eq) and 2, 2,
2-trifluoroethyl trifluoromethanesulfonate (6.46 g, 27.8 mmol, 1.1
eq) in DMF (30 mL) was added Cs.sub.2CO.sub.3 (12.4 g, 37.9 mmol,
1.5 eq). The reaction was stirred at 60-70.degree. C. for 16 hours
to give a light yellow suspension. TLC showed the reaction was
completed. The mixture was partitioned between water (100 mL) and
EtOAc (100 mL). The organic layer was washed with brine (100
mL.times.3), dried over anhydrous Na.sub.2SO.sub.4, filtered and
concentrated under reduced pressure to give A68 (6.1 g, crude) as a
white powder.
General Procedure for Synthesis A69
[0257] To a solution of A68 (1 g, 4.16 mmol, 1 eq) in THF (5 mL)
was added LDA (2 M, 2.29 mL, 1.1 eq) at -78.degree. C. under N2.
The reaction mixture was stirred for 5 min, and a solution of
I.sub.2 (1.16 g, 4.58 mmol, 1.1 eq) in THF (5 mL) was added. The
reaction was further stirred for 30 min at -78.degree. C., then
warmed to 20.degree. C. for 17 hours to give a yellow mixture. LCMS
showed most of starting material was still remained. The mixture
was quenched with water (5 mL), and 1 N HCl (1 mL) was added. The
mixture was partitioned between EtOAc (80 mL) and H.sub.2O (80 mL).
The organic layer was washed with saturated brine (80 mL.times.3),
dried over anhydrous Na.sub.2SO.sub.4, filtered, and concentrated
under reduced pressure to give a residue. The residue was purified
by combi flash (PE/EtOAc=I/O to 10/1) to give A69 (70 mg, 4.6%
yield) as yellow gum
General Procedure for Synthesis A70
[0258] To a solution of A69 (170 mg, 0.464 mmol, 1 eq) in THF (3
mL) was added LiOH (22 mg, 0.928 mmol, 2 eq) in H.sub.2O (1 mL).
The reaction was stirred at 20.degree. C. for 1.5 hours to give a
yellow mixture. LCMS showed the starting material was not consumed
completely. LiOH (20 mg) was added. The reaction was stirred at
20.degree. C. for 17 hours to give a yellow mixture. LCMS showed
the starting material was not consumed completely. LiOH (20 mg) was
added. The reaction was stirred at 40.degree. C. for 5 hours to
give a yellow mixture. TLC showed the reaction was completed. The
reaction was adjusted to pH=4 with 1 N HCl at 0-10.degree. C.,
extracted with DCM (30 mL.times.3). The combined organic layer was
dried over anhydrous Na.sub.2SO.sub.4, filtered and concentrated
under reduced pressure to give A70 (130 mg, 79.5% yield) as a
yellow gum
General Procedure for Synthesis C3
[0259] To a mixture of A70 (120 mg, 0.341 mmol, 1 eq) and B1-1 (101
mg, 0.341 mmol, 1 eq) in pyridine (2 mL) was added EDCI (131 mg,
0.682 mmol, 2 eq). The reaction was stirred at 20.degree. C. for 17
hours to give a yellow mixture. LCMS showed the reaction was
completed. The reaction mixture was concentrated under reduced
pressure to give a residue. The residue was partitioned between
EtOAc (20 mL) and H.sub.2O (20 mL). The aqueous layer was extracted
with EtOAc (20 mL.times.2). The combined organic layer was washed
with brine (30 mL), dried over anhydrous Na.sub.2SO.sub.4, filtered
and concentrated under reduced pressure to give a yellow gum. The
crude product was purified by prep-TLC (EtOAc/PE=1/3 first, then
DCM/MeOH=30:1) to give C3 (150 mg, 69.7% yield, 100% purity) as
off-white powder.
General Procedure for Synthesis 179
[0260] To a mixture of C3 (130 mg, 0.206 mmol, 1 eq), D1-3 (28 mg,
0.329 mmol, 1.6 eq), P(cy).sub.3 (8 mg, 0.021 mmol, 0.1 eq) and
K.sub.3PO.sub.4 (153 mg, 0.721 mmol, 3.5 eq) in toluene (2 mL) and
H.sub.2O (0.2 mL) was added Pd(OAc).sub.2 (5 mg, 0.021 mmol, 0.1
eq). The reaction was stirred at 100.degree. C. for 2 hours under
N.sub.2 to give a yellow mixture. LCMS showed the starting material
was not consumed completely. The reaction was stirred at
100.degree. C. for 2 hours under N.sub.2 to give a yellow mixture.
LCMS showed the starting material was not consumed completely.
cyclopropylboronic acid (30 mg), and K.sub.3PO.sub.4 (150 mg) were
added into the reaction mixture. The reaction was stirred at
100.degree. C. for 1 hour under N.sub.2 to give a yellow mixture.
LCMS showed the starting material was not consumed completely. The
mixture was partitioned between EtOAc (30 mL) and H.sub.2O (30 mL).
The aqueous layer was extracted with EtOAc (20 mL.times.2). The
combined organic layer was washed with brine (20 mL), dried over
anhydrous Na.sub.2SO.sub.4, filtered, and concentrated under
reduced pressure to give a residue. The crude product was purified
by prep-TLC (DCM/MeOH=40/1) to give yellow gum. LCMS showed impure.
The yellow gum was purified by prep-HPLC (column: Waters Xbridge
C18 150*50 mm*10 um; mobile phase: [water (0.04%
NH.sub.3H.sub.2O+10 mM NH.sub.4HCO.sub.3)-ACN]; B %: 50%-80%, 11
min). LCMS showed impure. The eluent was concentrated under reduced
pressure to give yellow oil. The oil was purified by prep-TLC
(EtOAc/PE=1/2) to give a yellow gum. LCMS showed impure. The yellow
gum was purified by prep-HPLC (column: Welch Xtimate C18 100*25
mm*3 um; mobile phase: [water (0.05% ammonia hydroxide v/v)-ACN]; B
%: 64%-64%, 12 min). The eluent was concentrated under reduced
pressure to give a residue. The residue was partitioned between
MeCN (5 mL) and H.sub.2O (5 mL) and lyophilized to give 179 (12.2
mg, 10.9% yield, 100% purity) as a white powder.
##STR00207##
General Procedure for Synthesis A71
[0261] To a solution of A6 (10 g, 42.0 mmol, 1 eq) and
3,4-dihydro-2H-pyran (10.6 g, 126 mmol, 11.5 mL, 3 eq) in THF (100
mL) was added p-TsOH (723 mg, 4.20 mmol, 0.1 eq). The resulting
mixture was stirred at 80.degree. C. for 3 hrs to give yellow
solution. TLC (PE:EtOAc=3:1) showed the reaction was completed. The
reaction mixture was concentrated under reduced pressure to give a
residue. Then diluted with H.sub.2O (100 mL) and extracted with
EtOAc (100 mL.times.2). The combined organic layers were washed
with brine (60 mL.times.2), dried over Na.sub.2SO.sub.4, filtered
and concentrated under reduced pressure to give a residue. The
residue was purified by column chromatography (SiO2, PE:EtOAc=3:1)
to give A71 (13 g, 94.6% yield, 98.5% purity) as a yellow oil.
General Procedure for Synthesis A72
[0262] To a solution of A71 (13 g, 40.3 mmol, 1 eq) in MeOH (60 mL)
and THF (60 mL) was added NaOH (3 M, 40.3 mL, 3 eq). The resulting
mixture was heated at 60.degree. C. and stirred for 1 hr to give
red solution. LCMS showed the reaction was completed. The reaction
mixture was concentrated under reduced pressure to give a residue.
Then diluted with H.sub.2O (50 mL) and and extracted with EtOAc
(100 mL.times.2). The water layers were adjust to pH-5 and
extracted with DCM (100 mL.times.2), washed with brine (30
mL.times.2), dried over Na.sub.2SO.sub.4, filtered and concentrated
under reduced pressure to give A72 (9.9 g, 83.4% yield, 100%
purity) as a yellow solid.
General Procedure for Synthesis C4
[0263] To a mixture of compound A72 (8.28 g, 28.2 mmol, 1.5 eq),
compound B1-1 (5.58 g, 18.8 mmol, 1 eq) in pyridine (15 mL) was
added EDCI (7.20 g, 37.5 mmol, 2 eq). The reaction was stirred at
20.degree. C. for 17 hours to give a yellow mixture. LCMS showed
the starting material was not consumed completely. The reaction
mixture was concentrated under reduced pressure to give a residue.
The residue was partitioned between EtOAc (200 mL) and H.sub.2O
(200 mL). The organic layer was washed with H.sub.2O (200
mL.times.2), brine (200 mL), dried over anhydrous Na.sub.2SO.sub.4,
filtered and concentrated under reduced pressure to give a yellow
gum. The crude product was purified by combi flash (EtOAc/PE=0/1 to
7/2) to give C4 (6.29 g, yield: 57%, purity: 98%) as yellow
gum.
General Procedure for Synthesis 16
[0264] To a solution of C4 (6.29 g, 11 mmol, 1 eq) in DCM (20 mL)
was added TFA (10 mL, 135 mmol, 12.3 eq) at 0.degree. C. The
reaction was stirred at 20.degree. C. for 17 hours to give a yellow
solution. LCMS showed the starting material was not consumed
completely. The mixture was concentrated under reduced pressure to
give a residue. The residue was dissolved in THF (20 mL). HCl (2 M,
11 mL, 2 eq) was added. The reaction was stirred at 20.degree. C.
for 2 hours to give a yellow solution. LCMS showed the starting
material was not consumed completely. HCl (2 M, 11 mL, 2 eq) was
added. The reaction was stirred at 20.degree. C. for 2 hours to
give a yellow solution. TLC showed the starting material was not
consumed completely. THF (25 mL) and HCl (2 M, 25 mL) were added.
The reaction was stirred at 50.degree. C. for 17 hours to give a
yellow solution. LCMS showed the starting material was not consumed
completely. The reaction was stirred at 55.degree. C. for 1.5 hours
to give a yellow solution. LCMS showed the starting material was
not consumed completely. After filtration, the filter cake was
washed with H.sub.2O (10 mL.times.2), and partitioned between
NaHCO.sub.3 (300 mL), DCM (300 mL), MeOH (50 mL). The aqueous layer
was extracted with DCM (300 mL). The combined organic layers were
dried over anhydrous Na.sub.2SO.sub.4, filtered and concentrated
under reduced pressure to give a yellow powder. The crude product
was triturated with EtOAc/PE (1/1, 40 mL) to give 16 (4.26 g,
yield: 79%, purity: 100%) as a white powder.
General Procedure for Synthesis C2
[0265] A method to prepare compounds of C2 is shown in Scheme 8.
The reaction of 16 and D1 is carried out in the presence of a
Cs.sub.2CO.sub.3 in a solvent like DMF or MeCN to give C2. In
addition, many of the D1 alkyl reagents are commercially
available.
##STR00208##
[0266] To a solution of 16 (20 mg, 0.409 mmol, 1 eq) and D1-11
(2.37 mg, 0.409 mmol, 2.86 uL, 1 eq) in CH.sub.3CN (2 mL) was added
Cs.sub.2CO.sub.3 (20 mg, 0.613 mmol, 1.5 eq). The resulting mixture
was stirred at 20.degree. C. for 16 hours to give yellow solution.
TLC (PE:EtOAc=0:1) showed little of the starting material was
remained and desired product was formed. The reaction mixture was
quenched by addition H.sub.2O (5 mL) and extracted with EtOAc (5
mL). The combined organic layers were washed with brine (3 mL),
dried over Na.sub.2SO.sub.4, filtered and concentrated under
reduced pressure to give a residue. The residue was purified by TLC
(PE:EtOAc=0:1), then dissolved in CH.sub.3CN (1 mL) and H.sub.2O (1
mL), lyophilized to afford 29 (14 mg, 62.6% yield, 100% purity) as
a white powder.
REFERENCES
[0267] The Role of TAM Family Receptors in Immune Cell Function:
Implications for Cancer Therapy. Paolino M, Penninger J M., Cancers
(Basel). 2016 Oct. 21; 8(10). pii: E97. [0268] Diversification of
TAM receptor tyrosine kinase function. Zagorska A, Traves P G, Lew
E D, Dransfield I, Lemke G., Nat Immunol. 2014 October;
15(10):920-8. [0269] TAM receptor tyrosine kinases as emerging
targets of innate immune checkpoint blockade for cancer therapy.
Akalu Y T, Rothlin C V, Ghosh S., Immunol Rev. 2017 March;
276(1):165-177. [0270] Ligand Activation of TAM Family
Receptors-Implications for Tumor Biology and Therapeutic Response.
Davra V, Kimani S G, Calianese D, Birge R B., Cancers (Basel). 2016
Nov. 29; 8(12). pii: E107. [0271] Development of monocytes,
macrophages, and dendritic cells. Geissmann F, Manz M G, Jung S,
Sieweke M H, Merad M, Ley K., Science. 2010 Feb. 5;
327(5966):656-61. [0272] CSF1/CSF1R blockade reprograms
tumor-infiltrating macrophages and improves response to T-cell
checkpoint immunotherapy in pancreatic cancer models. Zhu Y,
Knolhoff B L, Meyer M A, Nywening T M, West B L, Luo J, Wang-Gillam
A, Goedegebuure S P, Linehan D C, DeNardo D G., Cancer Res. 2014
Sep. 15; 74(18):5057-69.
[0273] The invention is now further exemplarily described by tables
1-7 which show activity data of selected compounds in binding
assays of example 1-2 (tables 1-2), in cellular ELISA assays of
examples 3-4 (tables 3 and 4), in cellular viability assays of
examples 4 and 5 (Tables 5 and 6), and comparison data of examples
6-7, Table 7, Example 8 and FIGS. 1 and 2; and the structure of
compounds 1-179 including .sup.1H-NMR-data is shown in table 8.
TABLE-US-00002 TABLE 1 Axl, Mer and CSF1R kinase binding activity #
cpds Axl Mer CSF1R 1 A A A 2 A A B 3 A A B 4 A A A 5 A A A 6 A A A
7 A A A 8 A A A 9 A A A 10 A A A 11 A A A 12 A A A 13 A A A 14 A A
A 15 A A n.d. 16 A A n.d. 17 A A n.d. 18 A A n.d. 19 A A n.d. 20 A
A n.d. 21 A A n.d. 22 A A n.d. 23 A A n.d. 24 A A n.d. 25 A A n.d.
26 A A n.d. 27 A A n.d. 28 A A n.d. 29 A A n.d. 30 A A n.d. 31 A A
n.d. 32 A A n.d. 33 A A n.d. 34 C B n.d. 35 C C n.d. 36 A A n.d. 37
B A n.d. 38 A A n.d. 39 A A n.d. 40 A A n.d. 41 A A n.d. 42 A A
n.d. 43 A A n.d. 44 A A n.d. 45 A A n.d. 46 A A n.d. 47 A A n.d. 48
A A n.d. 49 A A n.d. 50 A A n.d. 51 A A n.d. 52 A A n.d. 53 A A
n.d. 54 A A n.d. 55 A A n.d. 56 B A n.d. 57 A A n.d. 58 A A n.d. 59
A A n.d. 60 A A n.d. 61 A A n.d. 62 A A n.d. 63 A A n.d. 64 A A
n.d. 65 A A n.d. 66 A A n.d. 67 A A n.d. 68 A A n.d. 69 A A n.d. 70
A A n.d. 71 A A n.d. 72 A A n.d. 73 B B n.d. 74 B B n.d. 75 B B
n.d. 76 A A n.d. 77 A A n.d. 78 A B n.d. 79 A A n.d. 80 A A n.d. 81
C B n.d. 82 C B n.d. 83 A A n.d. 84 A A n.d. 85 A A n.d. 86 B B
n.d. 87 A A n.d. 88 B B n.d. 89 A A n.d. 90 A A n.d. 91 A A n.d. 92
A A n.d. 93 A A n.d. 94 A A n.d. 95 A A n.d. 96 A A n.d. 97 A A
n.d. 98 B A n.d. 99 A A n.d. 100 A A n.d. 101 A A n.d. 102 A A n.d.
103 A A n.d. 104 A A n.d. 105 A A n.d. 106 A A n.d. 107 A A n.d.
108 A A n.d. 109 A A n.d. 110 A A n.d. 111 A A n.d. 112 A A n.d.
113 A A n.d. 114 A A n.d. 115 A A n.d. 116 A A n.d. 117 A A n.d.
118 A A n.d. 119 B B n.d. 120 A A n.d. 121 A A n.d. 122 B B n.d.
123 A A n.d. 124 A A n.d. 125 C C n.d. 126 A A n.d. 127 C C n.d.
128 A A n.d. 129 B B n.d. 130 A A n.d. 131 A A n.d. 132 C C n.d.
133 B B n.d. 134 A A n.d. 135 C C n.d. 136 A A n.d. 137 C C n.d.
138 A A n.d. 139 A A n.d. 140 C B n.d. 141 C C n.d. 142 A A n.d.
143 A A n.d. 144 B B n.d. 145 A A n.d. 146 A A n.d. 147 A A n.d.
148 A A n.d. 149 A A n.d. 150 A A n.d. 151 A A n.d. 152 A A n.d.
153 A A n.d. 154 A A n.d. 155 A B n.d. 156 B B n.d. 157 B B n.d.
158 A A n.d. 159 C C n.d. 160 C C n.d. 161 A A n.d. 162 A A n.d.
163 A A n.d. 164 A A n.d. 165 B C n.d. 166 A A n.d. 167 A A n.d.
168 A A n.d. 169 A A n.d. 170 A A n.d. 171 A B n.d. 172 C C n.d.
173 A A n.d. 174 C C n.d. 175 A A n.d. 176 B B n.d. 177 A B n.d.
178 A A n.d. 179 B B n.d. Activity range: A indicates < 0.1 uM,
B indicates 0.1 .ltoreq. Kd < 0.5 uM, C indicates .gtoreq. 0.5
uM; n.d. = not determined
TABLE-US-00003 TABLE 2 CSF1R kinase binding activity (%, @ 0.1 uM)
# cpds Range Percent (%) 1 A 96 2 A 89 3 B 71 4 A 98 5 A 97 6 A 97
7 A 97 8 A 94 9 A 92 10 A 98 11 A 88 12 A 98 13 A 98 14 A 98 15 A
87 16 A 99 17 A 97 18 A 97 19 A 97 20 A 99 21 A 95 22 A 93 23 A 94
24 A 96 25 A 87 26 A 94 27 B 58 28 A 90 29 A 99 30 A 99 31 A 95 32
A 95 33 A 98 35 C 6 36 A 95 37 A 97 38 A 87 39 A 97 40 A 94 41 A 97
42 A 95 43 A 94 44 A 82 45 A 93 46 A 96 47 A 91 48 A 82 49 B 66 50
B 80 51 A 82 52 B 71 53 B 71 54 A 81 55 B 60 56 C 0 57 A 97 58 B 76
59 A 82 60 B 75 61 B 76 62 B 54 63 A 96 64 A 95 65 A 96 66 A 96 67
A 96 68 A 98 69 A 89 70 B 79 71 A 88 72 A 96 73 A 97 74 C 39 75 B
57 76 A 94 77 B 76 78 C 11 79 C 12 80 B 64 81 C 46 82 C 35 83 A 97
84 A 97 85 B 78 86 C 11 87 B 71 88 C 21 89 B 56 90 A 95 91 A 99 92
A 100 93 A 89 94 B 65 95 B 61 96 B 57 97 B 56 98 B 60 99 A 94 100 C
36 101 C 45 102 A 87 103 C 29 104 C 46 105 A 91 106 A 95 107 A 90
108 A 96 109 A 98 110 A 98 111 A 89 112 A 97 113 A 82 114 C 27 115
A 94 116 A 93 117 A 94 118 A 93 119 C 9 120 A 95 121 A 98 122 C 19
123 B 72 124 A 95 125 C 38 126 B 78 127 C 38 128 A 94 129 C 24 130
C 41 131 C 21 132 C 16 133 C 9 134 C 18 135 C 4 136 C 23 137 C 6
138 C 9 139 C 37 140 C 38 141 C 23 142 A 92 143 A 95 144 B 73 145 A
94 146 C 43 147 A 85 148 B 60 149 B 76 150 B 74 151 A 97 152 A 96
153 A 95 154 C 47 155 C 22 156 C 49 157 A 81 158 A 91 159 B 64 160
C 25 161 C 14 162 A 91 163 A 93 164 B 73 165 C 5 166 A 84 167 C 0
168 A 95 169 A 93 170 A 90 171 B 60 172 C 36 173 B 69 174 C 25 175
A 97 176 C 40 Activity range: A indicates .gtoreq. 80%, B indicates
80 > % inhibition 50%, C indicates < 50%
TABLE-US-00004 TABLE 3 Cellular Axl activity by H1299 Elisa # cpds
Axl 1 A 2 A 3 A 4 A 5 A 6 A 7 A 8 A 9 A 10 A 11 A 12 A 13 A 14 A 15
C 16 A 17 A 18 A 19 A 20 A 21 A 22 A 23 A 24 A 25 A 26 A 27 A 28 A
29 A 30 A 31 A 32 A 33 A 34 C 35 C 36 A 37 C 38 A 39 A 40 A 41 A 42
A 43 A 44 A 45 B 46 A 47 A 48 A 49 A 50 A 51 A 52 A 53 A 54 A 55 A
56 C 57 A 58 A 59 A 60 A 61 A 62 A 63 A 64 A 65 A 66 A 67 A 68 A 69
A 70 A 71 A 72 C 73 C 74 A 75 A 76 A 77 A 78 A 79 A 80 A 81 C 82 C
83 A 84 A 85 A 86 A 87 A 88 B 89 A 90 A 91 A 92 A 93 A 94 A 95 A 96
A 97 A 98 C 99 A 100 B 101 A 102 C 103 A 104 B 105 A 106 A 107 A
108 A 109 A 110 A 111 A 112 A 113 A 114 A 115 A 116 A 117 B 118 A
119 C 120 B 121 A 122 C 123 A 124 A 125 C 126 A 127 C 128 A 129 C
130 C 131 A 132 C 133 C 134 A 135 C 136 C 137 C 138 A 139 A 140 C
141 C 142 A 143 A 144 C 145 A 146 A 147 A 148 A 149 A 150 A 151 A
152 B 153 A 154 B 155 A 156 C 157 C 158 A 159 C 160 C 161 A 162 A
163 A 164 A 165 A 166 A 167 A 168 A 169 C 170 A 171 A 172 C 173 A
174 C 175 A 176 A 177 A 178 A 179 A Activity range: A indicates
< 0.5 uM, B indicates 0.5 .ltoreq. IC50 < 1 uM, C indicates
.gtoreq. 1 uM
TABLE-US-00005 TABLE 4 Cellular CSF1R activity by THP-1 Elisa assay
# cpds CSF1R 1 A 2 A 3 A 4 A 5 A 6 A 7 A 8 A 9 A 10 A 11 B 12 A 13
A 14 A 15 C 16 A 17 A 18 A 19 A 20 A 21 A 22 A 23 A 24 A 25 B 26 A
27 C 28 B 29 A 30 A 31 A 32 A 33 A 34 C 35 C 36 A 37 A 38 B 39 A 40
A 41 A 42 A 43 A 44 A 45 A 46 B 47 B 48 B 49 C 50 B 51 A 52 C 53 B
54 C 55 C 56 A 57 B 58 C 59 A 60 B 61 C 62 C 63 B 64 A 65 A 66 A 67
A 68 A 69 A 70 B 71 A 72 A 73 A 74 C 75 C 76 C 77 C 78 C 79 C 80 C
81 C 82 C 83 A 84 B 85 B 86 C 87 B 88 C 89 B 90 A 91 A 92 A 93 A 94
C 95 C 96 C 97 C 98 C 99 B 100 C 101 C 102 A 103 C 104 C 105 A 106
A 107 A 108 A 109 A 110 A 111 B 112 B 113 A 114 C 115 B 116 B 117 A
118 B 119 C 120 B 121 A 122 C 123 C 124 B 125 C 126 C 127 C 128 A
129 C 130 C 131 C 132 C 133 C 134 C 135 C 136 C 137 C 138 C 139 C
140 C 141 C 142 A 143 A 144 C 145 A 146 C 147 B 148 C 149 C 150 C
151 A 152 A 153 A 154 C 155 C 156 C 157 B 158 A 159 C 160 C 161 C
162 A 163 B 164 C 165 C 166 C 167 C 168 A 169 A 170 A 171 C 172 C
173 C 174 C 175 A 176 C 177 C 178 A 179 A Activity range: A
indicates < 0.5 uM, B indicates 0.5 .ltoreq. IC50 < 1 uM, C
indicates .gtoreq. 1 uM
TABLE-US-00006 TABLE 5 Cellular CSF1R activity by M-NFS-60
viability assay # cpds CSF1R 1 A 2 A 3 C 4 A 5 A 6 A 7 A 8 A 9 A 10
A 11 A 12 A 13 A 14 A 15 B 16 A 17 A 18 A 19 A 20 A 21 A 22 A 23 A
24 A 25 B 26 B 27 B 28 B 29 A 30 A 31 A 32 B 33 A 34 B 35 B 36 A 37
A 38 A 39 A 40 A 41 A 42 A 43 A 44 A 45 B 46 B 47 A 48 A 49 B 50 B
51 A 52 A 53 A 54 A 55 B 56 A 57 A 58 B 59 A 60 B 61 B 62 B 63 B 64
B 65 B 66 B 67 B 68 B 69 B 70 B 71 B 72 B 73 B 74 B 75 B 76 B 77 B
78 B 79 B 80 B 81 B 82 B 83 B 84 B 85 B 86 B 87 B 88 B 89 B 90 A 91
A 92 A 93 A 94 B 95 B 96 B 97 B 98 B 99 B 100 B 101 B 102 B 103 B
104 C 105 A 106 A 107 B 108 A 109 A 110 A 111 B 112 B 113 B 114 B
115 B 116 B 117 A 118 B 119 B 120 B 121 A 122 B 123 B 124 B 125 C
126 B 127 C 128 B 129 C 130 B 131 B 132 C 133 B 134 C 135 C 136 B
137 B 138 B 139 B 140 B 141 C 142 B 143 B 144 C 145 A 146 B 147 B
148 B 149 B 150 B 151 A 152 A 153 A 154 B 155 B 156 B 157 B 158 B
159 C 160 C 161 B 162 B 163 B 164 C 165 B 166 B 167 B 168 A 169 A
170 A 171 C 172 C 173 B 174 C 175 A 176 B 177 B 178 A 179 A
Activity range: A indicates < 1.0 uM, B indicates 1.0 .ltoreq.
IC50 < 10 uM, C indicates .gtoreq. 10 uM
TABLE-US-00007 TABLE 6 Cellular Axl and Mer activity by Ba/F3 assay
# cpds Axl Mer CSF1R 4 A A A 5 A A A 7 A A A 10 A A A 12 A A A 15 A
B B 16 A A A 17 A A A 18 A A A 29 A A A 30 A A A 35 C C B Activity
range: A indicates < 1.0 uM, B indicates 1.0 .ltoreq. IC50 <
10 uM, C indicates .gtoreq. 10 uM
TABLE-US-00008 TABLE 7 Comparison data by Binding and Cellular
assay Binding activity Cellular activity Axl Mer Axl (H1299) # Cpds
(Kd, nM) (Kd, nM) (IC50, nM) Compound 22* 104 73 >3000 Compound
16** 19 12 44 Compound 27* 64 11 190 Compound 4** 1.1 1.6 3.4
Compound 48* 19 3.7 183 Compound 92** 1.4 1.3 3.9 Compound 64* 60
3.2 337 Compound 10** 3.2 0.2 5.2 *: Compounds of WO2016/166250 **:
Compounds of present invention
TABLE-US-00009 TABLE 8 Summarizes compounds 1-179 in terms of their
structures and corresponding characteristics. # cpd Structure
Characterization Data 1 ##STR00209## yellow powder; .sup.1H-NMR
(DMSO-d6, 400 MHz): .delta. 9.59 (1H, s), 8.50 (1H, d, J = 5.2 Hz),
8.40 (1H, d, J = 2.8 Hz), 8.31 (1H, d, J = 9.2 Hz), 7.87 (1H, dd, J
= 8.8, 2.4 Hz), 7.54 (1H, s), 7.42 (1H, s), 6.55 (1H, d, J = 5.2
Hz), 4.73 (2H, q, J = 9.2 Hz), 4.15 (2H, q, J = 7.2 Hz), 3.95 (3H,
s), 3.94 (3H, s), 2.24 (3H, s), 1.38 (3H, t, J = 7.2 Hz); LCMS:
99.1%, MS (ESI): m/z 532.1 [M + H]+. 2 ##STR00210## white powder;
.sup.1H-NMR (DMSO-d6, 400 MHz): .delta. 9.60 (1H, s), 8.51 (1H, d,
J = 5.2 Hz), 8.41 (1H, d, J = 2.8 Hz), 8.32 (1H, d, J = 9.2 Hz),
7.87 (1H, dd, J = 9.2, 2.8 Hz), 7.54 (1H, s), 7.42 (1H, s), 6.55
(1H, d, J = 5.2 Hz), 4.73 (2H, q, J = 9.2 Hz), 4.54-4.65 (1H, m),
3.96 (3H, s), 3.95 (3H, s), 2.25 (3H, s), 1.43 (6H, d, J = 6.4 Hz);
LCMS: 96.9%, MS (ESI): m/z 546.1 [M + H]+. 3 ##STR00211## white
powder; .sup.1H-NMR (DMSO-d6, 400 MHz): .delta. 10.25 (1H, brs),
8.49 (1H, d, J = 5.2 Hz), 8.43 (1H, d, J = 3.2 Hz), 8.28 (1H, 4
##STR00212## white powder; .sup.1H-NMR (DMSO-d6, 400 MHz): 9.53
(1H, brs), 8.50 (1H, d, J = 5.2 Hz), 8.40 (1H, d, J = 3.2 Hz), 8.32
(1H, d, J = 8.8 Hz), 7.99 (1H, s), 7.86 (1H, dd, J = 8.8, 2.8 Hz),
7.54 (1H, s), 7.42 (1H, s), 6.55 (1H, d, J = 5.2 Hz), 4.78 (2H, q,
J = 8.8 Hz), 4.18 (2H, q, J = 7.2 Hz), 3.96 (3H, s), 3.95 (6H, s),
1.43 (3H, t, J = 7.2 Hz); LCMS: 100%, MS (ESI): m/z 518.1 [M + H]+.
5 ##STR00213## white powder; .sup.1H-NMR (DMSO-d6, 400 MHz):
.delta. 9.58 (1H, brs), 8.50 (1H, d, J = 5.2 Hz), 8.40 (1H, d, J =
2.8 Hz), 8.31 (1H, d, J = 8.8 Hz), 8.04 (1H, s), 7.86 (1H, dd, J =
8.8, 2.8 Hz), 7.54 (1H, s), 7.42 (1H, s), 6.55 (1H, d, J = 5.2 Hz),
4.76 (2H, q, J = 8.8 Hz), 3.95 (3H, s), 3.94 (3H, s), 3.78-3.87
(1H, m), 1.11- 1.18 (2H, m), 0.99-1.07 (2H, m); LCMS: 98.4%, MS
(ESI): m/z 530.1 [M + H]+. 6 ##STR00214## white powder; .sup.1H-NMR
(DMSO-d6, 400 MHz): .delta. 9.58 (1H, brs), 8.49 (1H, d, J = 5.2
Hz), 8.39 (1H, d, J = 3.2 Hz), 8.36 (1H, d, J = 8.8 Hz), 7.82-7.89
(2H, m), 7.54 (1 H, s), 7.41 (1H, s), 6.55 (1H, d, J = 5.2 Hz),
4.35-4.46 (1H, m), 4.15 (2H, q, J = 7.2 Hz), 3.95 (3H, s), 3.94
(3H, s), 1.41 (3H, t, J = 7.2 Hz), 1.36 (6 H, d, J = 6.0 Hz); LCMS:
100%, MS (ESI): m/z 478.1 [M + H]+. 7 ##STR00215## white powder;
.sup.1H-NMR (DMSO-d6, 400 MHz): 9.51 (1H, brs), 8.51 (1H, d, J =
5.2 Hz), 8.40 (1H, d, J = 2.8 Hz), 8.32 (1H, d, J = 9.2 Hz), 7.97
(1H, s), 7.87 (1H, dd, J = 9.2, 2.4 Hz), 7.54 (1H, s), 7.42 (1H, s)
6.55 (1H, d, J = 5.2 Hz), 4.78 (2H, q, J = 8.8 Hz), 4.11 (2H, t, J
= 6.4 Hz), 3.95 (3H, s), 3.94 (3H, s), 1.81- 1.86 (2H, m), 0.86
(3H, t, J = 7.6 Hz); LCMS: 100%, MS (ESI): m/z 532.1 [M + H]+. 8
##STR00216## white powder; .sup.1H-NMR (DMSO-d6, 400 MHz): 9.59
(1H, brs), 8.49 (1H, d, J = 5.2 Hz), 8.34-8.39 (2H, m), 7.83-7.88
(2H, m), 7.53 (1H, s), 7.41 (1H, s), 6.55 (1H, d, J = 5.2 Hz), 4.15
(2H, q, J = 7.2 Hz), 4.03 (2H, d, J = 6.8 Hz), 3.95 (3H, s), 3.94
(3H, s), 2.72-2.84 (1H, m), 2.05-2.11 (2H, m), 1.87-1.98 (4H, m),
1.41 (3H, t, J = 7.2 Hz); LCMS: 100%, MS (ESI): m/z 504.1 [M + H]+.
9 ##STR00217## white powder; .sup.1H-NMR (DMSO-d6, 400 MHz): 9.56
(1H, brs), 8.48 (1H, d, J = 5.2 Hz), 8.38 (1H, d, J = 2.8 Hz), 8.30
(1H, d, J = 9.2 Hz), 7.96 (1H, s), 7.85 (1H, dd, J = 9.2, 2.8 Hz),
7.52 (1H, s), 7.40 (1H, s), 6.53 (1H, d, J = 4.8 Hz), 4.75 (2H, q,
J = 8.8 Hz), 4.20-4.35 (2H, m), 3.94 (3H, s), 3.93 (3H, s),
2.65-2.90 (2H, m), 2.10-2.35 (6H, m); LCMS: 100%, MS (ESI): m/z
583.1 [M + Na]+. 10 ##STR00218## white powder; .sup.1H-NMR
(DMSO-d6, 400 MHz): 9.54 (1H, brs), 8.50 (1H, d, J = 5.2 Hz), 8.40
(1H, d, J = 2.8 Hz), 8.32 (1H, d, J = 9.2 Hz), 7.95 (1H, s), 7.87
(1H, dd, J = 8.8, 2.8 Hz), 7.54 (1H, s), 7.42 (1H, s), 6.56 (2H, d,
J = 4.8 Hz), 4.79 (2H, q, J = 8.8 Hz), 4.31 (2H, t, J = 5.2 Hz),
3.95 (3H, s), 3.94 (3H, s), 3.75 (2H, d, J = 5.2 Hz), 3.26 (3H, s);
LCMS: 100%, MS (ESI): m/z 548.1 [M + H]+. 11 ##STR00219## white
powder; .sup.1H-NMR (DMSO-d6, 400 MHz): .delta. 9.57 (1H, brs),
8.50 (1H, d, J = 5.2 Hz), 8.40 (1H, d, J = 3.2 Hz), 8.32 (1H, d, J
= 9.2 Hz), 7.94 (1H, s), 7.86 (1H, d, J = 8.8, 2.8 Hz), 7.53 (1H,
s), 7.42 (1H, s), 6.55 (1H, d, J = 5.2 Hz), 4.76 (2H, q, J = 8 8
Hz), 4.19 (2H, q, J = 6.0 Hz),3.95 (3H, s), 3.94 (3H, s), 2.90 (2H,
q, J = 6.0 Hz), 2.29 (3H, s); LCMS: 100%, MS (ESI): m/z 569.1 [M +
Na]+ 12 ##STR00220## white powder; .sup.1H-NMR (DMSO-d6, 400 MHz):
.delta. 9.48 (1H, brs), 8.50 (1H, d, J = 5.2 Hz), 8.38 (1H, d, J =
3.2 Hz), 8.31 (1H, d, J = 8.8 Hz), 7.91 (1H, s), 7.85 (1H, dd, J =
9.2, 2.8 Hz), 7.53 (1H, s), 7.41 (1H, s), 6.20-6.70 (2H, m),
4.25-4.44 (2H, m), 4.08 (2H, t, J = 7.2 Hz), 3.94 (3H, s), 3.93
(3H, s), 1.75-1.90 (2H, m), 0.85 (2H, t, J = 7.6 Hz); LCMS: 100%,
MS (ESI): m/z 514.1 [M + H]+. 13 ##STR00221## white powder;
.sup.1H-NMR (400 MHz, DMSO-d6) .delta. 9.61 (1H, brs), 8.50 (1H, d,
J = 4.8 Hz), 8.40 (1H, d, J = 2.4 Hz), 8.31 (1H, d, J = 9.6 Hz),
8.02 (1H, s), 7.87 (1H, dd, J = 8.8, 2.8 Hz), 7.54 (1H, s), 7.42
(1H, s), 6.56 (2H, d, J = 5.6 Hz), 5.02-5.08 (1H, m), 4.79 (2H, q,
J = 8.8 Hz), 3.90-4.05 (9H, m), 3.81-3.83 (1H, m), 2.30-2.38 (2H,
m); LCMS: 100%, MS (ESI): m/z 560.1 [M + H]+. 14 ##STR00222## white
powder; .sup.1H-NMR (DMSO-d6, 400 MHz): .delta. 9.52 (1H, s), 8.49
(1H, d, J = 5.2 Hz), 8.39 (1H, d, J = 2.8 Hz), 8.34 (1H, d, J = 9.2
Hz), 7.84-7.89 (2H, m), 7.54 (1H, s), 7.41 (1H, s), 6.55 (1H, d, J
= 5.2 Hz), 4.69-4.85 (2H, m), 4.25-4.36 (2H, m), 4.09 (2H, t, J =
6.8 Hz), 3.95 (6H, d, J = 2.8 Hz), 1.79-1.88 (2H, m), 0.86 (3H, t,
J = 7.6 Hz); LCMS: 96.7%, MS (ESI): m/z 496.1 [M + H]+. 15
##STR00223## white powder; .sup.1H-NMR (DMSO-d6, 400 MHz): .delta.
9.55 (1H, brs), 8.49 (1H, d, J = 5.2 Hz), 8.31-8.41 (2H, m),
7.80-7.90 (2H, m), 7.54 (1H, S), 7.41 (1H, s), 6.55 (1H, d, J = 5.2
Hz), 4.68 (1H, d, J = 4.8 Hz), 4.59 (1H, t, J = 5.6 Hz), 4.14 (2H,
t, J = 6.0 Hz), 4.08 (2H, t, J = 6.8 Hz), 3.95 (3H, s), 3.94 (3H,
s), 3.64-3.73 (1H, m), 3.35-3.44 (2H, m), 1.95-2.07 (1H, m),
1.79-1.89 (2H, m), 1.64-1.77 (1H, m), 0.86 (3H, t, J = 7.2 Hz);
LCMS: 100%, MS (ESI): m/z 538.2 [M + H]+. 16 ##STR00224## white
powder; .sup.1H-NMR (DMSO-d6, 400 MHz) .delta. 13.84 (1H, brs),
13.40 (1H, brs), 9.61 (1H, brs), 9.29 (1H, brs), 8.50 (1H, d, J =
5.2 Hz), 8.40 (1H, s), 8.33 (1H, d, J = 8.8 Hz), 7.93 (1H, s), 7.87
(1H, d, J = 8.8 Hz), 7.77 (1H, s), 7.54 (1H, s), 7.42 (1H, s), 6.55
(1H, d, J = 5.2 Hz), 5.01 (1H, d, J = 8.0 Hz), 4.77 (1H, q, J = 8.8
Hz), 3.95 (3H, s), 3.94 (3H, s); LCMS: 100%, MS (ESI): m/z 490.0 [M
+ H]+. 17 ##STR00225## white solid; .sup.1H-NMR (CDCl3, 400 MHz)
.delta. 9.30 (1H, s), 8.48- 8.53 (2H, m), 8.27-8.29 (m, 1H), 7.62
(1H, dd, J = 8.8 Hz), 7.56 (1H, s), 7.43 (1H, s), 7.41 (1H, s),
6.46 (1H, d, J = 6.4 Hz), 4.49 (2H, q, J = 8.4 Hz), 4.28-4.34 (1H,
m), 4.16-4.20 (1H, m), 4.07 (6H, s), 3.88-4.04 (1H, m), 1.29 (3H,
d, J = 6.4 Hz); LCMS: 100.0%, MS (ESI): m/z 548.1 [M + H]+. 18
##STR00226## off-white powder; .sup.1H-NMR (DMSO-d6, 400 MHz)
.delta. 9.54 (1H, s), 8.50 (1H, d, J = 5.2 Hz), 8.40 (1H, d, J =
3.2 Hz), 8.32 (1H, d, J = 8.8 Hz), 7.96 (1H, s), 7.86 (1H, dd, J =
9.2, 3.2 Hz), 7.54 (1H, s), 7.42 (1H, s), 6.55 (1H, d, J = 5.6 Hz),
4.77 (2H, q, J = 9.2 Hz), 4.67 (1H, t, J = 4.8 Hz), 4.20 (2H, t, J
= 6.8 Hz), 3.95 (3H, s), 3.94 (3H, s), 3.42 (2H, q, J = 5.6 Hz),
1.92-2.05 (2H, m); LCMS: 100%, MS (ESI): m/z 548.1 [M + H]+. 19
##STR00227## yellowish powder; .sup.1H-NMR (CDCl3, 400 MHz) .delta.
9.29 (1H, s), 8.53 (1H, d, J = 5.2 Hz), 8.48 (1H, d, J = 9.2 Hz),
8.27 (1H, d, J = 2.4 Hz), 7.61 (1H, dd, J = 8.8 Hz, 2.8 Hz), 7.56
(1H, s), 7.45 (1H, s), 7.38 (1H, s), 6.47 (1H, d, J = 5.2 Hz), 4.94
(2H, s), 4.57 (2H, q, J = 8.4 Hz), 4.07 (6H, s), 2.25 (3H, s);
LCMS: 100.0%, MS (ESI): m/z 546.0 [M + H]+. 20 ##STR00228## white
powder; .sup.1H-NMR (DMSO-d6, 400 MHz) .delta. 10.33 (1H, s), 8.54
(1H, s), 8.51 (1H, d, J = 5.2 Hz), 8.44 (1H, d, J = 2.8 Hz), 8.31
(1H, d, J = 8.8 Hz), 7.89 (1H, dd, J = 9.2, 3.2 Hz), 7.54 (1H, s),
7.42 (1H, s), 6.57 (1H, d, J = 5.2 Hz), 4.89 (2H, q, J = 8.4 Hz),
3.95 (3H, s), 3.94 (3H, s), 3.22 (2H, q, J = 7.2 Hz), 1.19- 1.21
(3H, m); LCMS: 97.6%, MS (ESI): m/z 546.1 [M + H]+. 21 ##STR00229##
white powder; .sup.1H-NMR (CDCl3, 400 MHz) .delta. 9.36 (1H, s),
8.53 (1H, d, J = 5.2 Hz), 8.49 (1H, d, J = 8.8 Hz), 8.29 (1H, d, J
= 2.8 Hz), 7.61 (1H, dd, J = 8.8, 2.8 Hz), 7.56 (1H, s), 7.45 (1H,
s), 7.31 (1H, s), 6.47 (1H, d, J = 5.2 Hz), 4.56 (2H, t, J = 8.4
Hz), 4.07 (6H, s), 3.90 (2H, d, J = 7.2 Hz), 2.21-2.27 (1H, m),
0.95 (6H, d, J = 6.8 Hz); LCMS: 100.0%, MS (ESI): m/z 546.1 [M +
H]+. 22 ##STR00230## white powder; 1H-NMR (CDCl3, 400 MHz) .delta.
9.37 (1H, s), 8.50-8.53 (2H, m), 8.29 (1H, d, J = 2.8 Hz), 7.61
(1H, dd, J = 9.2, 2.8 Hz), 7.57 (1H, s), 7.45-7.46 (2H, m), 6.47
(1H, d, J = 5.6 Hz), 4.56 (2H, q, J = 8.4 Hz), 4.07-4.08 (6H, m),
1.62 (6H, s); LCMS: 100.0%, MS (ESI): m/z 546.1 [M + H]+. 23
##STR00231## white powder; .sup.1H-NMR (CDCl3, 400 MHz) .delta.
9.24 (1H, s), 8.53 (1H, d, J = 5.2 Hz), 8.44 (1H, d, J = 9.2 Hz),
8.27 (1H, d, J = 2.4 Hz), 7.59 (1H, dd, J = 9.2, 2.8 Hz), 7.56 (1H,
s), 7.45 (1H, s), 6.97 (1H, s), 6.47 (2H, d, J = 5.6 Hz), 5.72 (1H,
t, J = 7.0 Hz), 4.07-4.08 (6H, m), 4.03 (2H, t, J = 7.0 Hz),
3.62-3.68 (2H, m), 1.87-1.94 (2H, m), 0.95 (2H, t, J = 7.6 Hz);
LCMS: 100.0%, MS (ESI): m/z 531.1 [M + H]+. 24 ##STR00232## white
powder; .sup.1H-NMR (DMSO-d6, 400 MHz): .delta. 9.58 (1H, s), 8.50
(1H, d, J = 5.2 Hz), 8.40 (1H, d, J = 3.2 Hz), 8.32 (1H, d, J = 9.2
Hz), 8.08 (1H, s), 7.87 (1H, dd, J = 9.2, 3.2 Hz), 7.54 (1H, s),
7.42 (1H, s), 6.56 (1H, d, J = 4.8 Hz), 4.82-4.91 (1H, m), 4.78
(2H, q, J = 8.8 Hz), 3.95 (3H, s), 3.95 (3H, s), 2.31-2.47 (4H, m),
1.75-1.89 (2H, m); LCMS: 100%, MS (ESI): m/z 544.1 [M + H]+. 25
##STR00233## white foam; .sup.1H-NMR (DMSO-d6, 400 MHz): .delta.
10.24 (1H, s), 8.48 (1H, d, J = 4.8 Hz), 8.03 (1H, dd, J = 13.6,
2.0 Hz), 7.92 (1H, s), 7.74 (1H, dd, J = 7.6, 2.0 Hz), 7.53 (1H,
s), 7.47-7.40 (2H, m), 6.46 (1H, d, J = 5.6 Hz), 4.70 (2H, q, J =
8.8 Hz), 4.10 (2H, t, J = 6.8 Hz), 3.95 (6H, s), 1.89-1.80 (2H, m),
0.87 (3H, t, J = 7.2 Hz); LCMS: 100.0%, MS (ESI): m/z 549.1 [M +
H]+. 26 ##STR00234## white powder; .sup.1H-NMR (DMSO-d6, 400 MHz):
.delta. 9.89 (1H, s), 8.44 (1H, d, J = 5.2 Hz), 7.91 (1H, s), 7.87
(1H, d, J = 2.4 Hz), 7.72 (1H, dd, J = 8.8, 2.4 Hz), 7.57 (1H, s),
7.39 (1H, s),7.16 (1H, d, J = 8.4 Hz), 6.29 (1H, d, J = 5.2 Hz),
4.70 (2H, q, J = 8.8 Hz), 4.09 (2H, t, J = 6.8 Hz), 3.95 (6H, s),
2.1 (3H, s), 1.89-1.80 (2H, m), 0.87 (3H, t, J = 7.2 Hz); LCMS:
100.0%, MS (ESI): m/z 545.1 [M + H]+. 27 ##STR00235## white powder;
.sup.1H-NMR (DMSO-d6, 400 MHz): .delta. 9.21 (1H, s), 8.48 (1H, d,
J = 5.2 Hz), 7.92 (1H, s), 7.81 (1H, d, J = 8.8 Hz), 7.50 (1H, s),
7.40 (1H, s), 7.20 (1H, d, J = 2.8 Hz), 7.12 (1H, dd, J = 8.4, 2.4
Hz), 6.50 (1H, d, J = 5.2 Hz), 4.74 (2H, q, J = 9.2 Hz), 4.08 (2H,
t, J = 6.8 Hz), 3.94 (3H, s), 3.93 (3H, s), 2.8 (3H, s), 1.89-1.78
(2H, m), 0.87 (3H, t, J = 7.2 Hz); LCMS: 100.0%, MS (ESI): m/z
545.1 [M + H]+. 28 ##STR00236## white powder; .sup.1H-NMR (DMSO-d6,
400 MHz): .delta. 9.4 (1H, s), 8.52 (1H, d, J = 5.2 Hz), 8.05 (1H,
t, J = 9.2 Hz), 7.94 (1H, s), 7.48 (1H, s), 7.42-7.35 (2H, m), 7.14
(2H, dd, J = 10.4, 1.6 Hz), 6.59 (1H, d, J = 5.2 Hz), 4.74 (2H, q,
J = 8.8 Hz), 4.09 (2H, t, J = 6.8 Hz), 3.93 (3H, s), 3.94 (3H, s),
1.9-1.76 (2H, m), 0.87 (3H, t, J = 7.2 Hz); LCMS: 100.0%, MS (ESI):
m/z 549.1 [M + H]+. 29 ##STR00237## (R), white powder; .sup.1H-NMR
(CDCl3, 400 MHz) .delta. 9.30 (1H, s), 8.49-8.51 (2H, m), 8.28 (1H,
d, J = 2.8 Hz), 7.61 (1H, dd, J = 9.2, 2.8 Hz), 7.56 (1H, s), 7.44
(1H, s), 7.42 (1H, s), 6.46 (1H, d, J = 5.2 Hz), 4.52 (1H, q, J =
8.0 Hz), 4.19-4.20 (1H, m), 4.17- 4.19 (1H, m), 4.07 (6H, s),
4.01-4.04 (1H, m), 1.28 (3H, d, J = 6.4 Hz); LCMS: 100.0%, MS
(ESI): m/z 548.1 [M + H]+. 30 ##STR00238## (S), white powder;
.sup.1H-NMR (CDCl3, 400 MHz) .delta. 9.30 (1H, s), 8.49-8.51 (2H,
m), 8.28 (1H, d, J = 2.8 Hz), 7.61 (1H, dd, J = 8.8, 2.8 Hz), 7.56
(1H, s), 7.45 (1H, s), 7.42 (1H, s), 6.46 (1H, d, J = 5.2 Hz), 4.51
(1H, q, J = 8.4 Hz), 4.19-4.20 (1H, m), 4.17- 4.19 (1H, m), 4.07
(6H, s), 4.02-4.04 (1H, m), 1.29 (3H, d, J = 6.4 Hz); LCMS: 100.0%,
MS (ESI): m/z 548.1 [M + H]+. 31 ##STR00239## yellow powder;
.sup.1H-NMR (CDCl3, 400 MHz): .delta. 9.34 (1H, s), 8.49-8.55 (2H,
m), 8.26 (1H, d, J = 2.8 Hz), 7.54-7.61 (2H, m), 7.46 (1H, s), 7.20
(1H, s), 6.47 (1H, d, J = 5.2 Hz), 4.28 (2H, t, J = 6.8 Hz),
4.03-4.11 (8H, m), 2.74 (2H, m), 1.93 (2H, m), 0.95 (3H, t, J = 7.6
Hz); LCMS: 100%, MS (ESI): m/z 546.1 [M + H] +. 32 ##STR00240##
yellow powder; .sup.1H-NMR (400 MHz, CDCl3): .delta. 9.69 (s, 1H),
8.80 (d, J = 9.6 Hz, 1H), 8.64 (d, J = 5.2 Hz, 1H), 7.39-7.46 (m,
3H), 7.31 (s, 1H), 6.92 (d, J = 5.2 Hz, 1 H), 4.51 (q, J = 8.4 Hz,
2H), 4.00-4.10 (m, 8H), 1.89-1.96 (m, 2H), 0.94 (t, J = 7.6 Hz,
3H); LCMS: 100%, MS (ESI): m/z 533.1 [M + H]+. 33 ##STR00241##
white powder; .sup.1H-NMR (400 MHz, DMSO-d6) .delta. ppm 9.51 (1H,
s), 8.50 (1H, d, J = 5.2 Hz), 8.39 (1H, d, J = 2.8 Hz), 8.31 (1H,
d, J = 8.8 Hz), 7.91 (1H, s), 7.86 (1H, dd, J = 8.8, 2.8 Hz), 7.54
(1H, s), 7.41 (1H, s), 6.55 (1H, d, J = 5.2 Hz), 4.77 (2H, q, J =
8.8 Hz), 3.94-3.97 (6H, m), 3.90 (3H, s); LCMS: 100%, MS (ESI): m/z
504.0 [M + H]+. 34 ##STR00242## white powder; .sup.1H-NMR (400 MHz,
CDCl3): .delta. 10.83 (1H, s), 8.47-8.63 (2H, m), 8.24 (1H, d, J =
2.4 Hz), 7.53-7.62 (2H, m), 7.45 (1H, s), 7.33 (1H, s), 6.48 (1H,
d, J = 5.2 Hz), 4.12 (2H, t, J = 7.2 Hz), 4.07 (3H, s), 4.06 (3H,
s), 3.92-4.03 (2H, m), 3.31- 3.41 (2H, m), 2.89 (2H, d, J = 7.2
Hz), 2.78 (3H, s), 1.91-1.99 (2H, m), 1.82-1.91 (1H, m), 1.73-1.82
(3H, m), 1.33-1.41 (2H, m), 0.95 (2H, t, J = 7.2 Hz); LCMS: 100%,
MS (ESI): m/z 561.2 [M + H]+. 35 ##STR00243## white powder;
.sup.1H-NMR (CDCl3, 400 MHz): .delta. 9.17 (1H, s), 8.31 (1H, s),
8.04 (1H, d, J = 11.6 Hz), 7.46-7.50 (2H, m), 7.36-7.40 (1H, m),
7.18 (1H, s), 6.34-6.36 (2H, m), 4.35-4.42 (1H, m), 4.11 (2H, t, J
= 7.2 Hz), 3.74 (3H, s), 1.89-1.99 (2H, m), 1.47 (6H, d, J = 6.0
Hz), 0.96 (3H, t, J = 7.2 Hz); LCMS: 100.0%, MS (ESI): m/z 563.1 [M
+ H]+.
36 ##STR00244## white solid; .sup.1H-NMR (CDCl3, 400 MHz): .delta.
9.28 (s, 1H), 8.45 (d, J = 4.2 Hz, 1H), 8.42 (d, J = 9.2 Hz, 1H),
8.21 (d, J = 2.4 Hz, 1H), 7.53 (dd, J = 3.2, 9.2 Hz, 1H), 7.49 (s,
1H), 7.40 (br.s, 1H), 7.24 (s, 1H), 6.40 (d, J = 5.2 Hz, 1H), 4.48
(q, J = 8.4 Hz, 2H), 4.03 (t, J = 7.2 Hz, 2H), 4.00 (s, 6H),
1.77-1.85 (m,2H), 1.24- 1.33 (m,2H), 0.90 (t, J = 7.6 Hz, 3H);
LCMS: 96.8%, MS (ESI): m/z 546.1 [M + H]+. 37 ##STR00245## white
solid; .sup.1H-NMR (CDCl3, 400 MHz): .delta. 8.56 (s, 1H), 8.39 (d,
J = 5.2 Hz, 1H), 7.63 (d, J = 2.4 Hz, 1H), 7.55 (s, 1H), 7.49 (dd,
J = 2.4. 8.8 Hz, 1H), 7.40 (s, 1H), 7.19 (s, 1H), 7.03 (d, J = 8.8
Hz, 1H), 6.25 (d, J = 5.2 Hz, 1H), 4.49 (q, J = 8.4 Hz, 2H), 4.00
(s, 3H), 3.99 (s, 3H), 3.85 (s, 3H), 2.13 (s, 3H); LCMS: 95.6%, MS
(ESI): m/z 517.2 [M + H]+. 38 ##STR00246## white solid; .sup.1H-NMR
(CDCl3, 400 MHz): .delta. 8.69 (s, 1H), 8.50 (d, J = 5.2 Hz, 1H),
7.78 (d, J = 8.8 Hz, 2H), 7.58 (s, 1H), 7.45 (s, 1H), 7.33 (s, 1H),
7.21 (d, J = 8.8 Hz, 2H), 6.49 (d, J = 5.2 Hz, 1H), 4.57 (q, J =
8.4 Hz, 2H), 4.07-4.10 (m, 8H), 1.91-1.97 (m, 2H), 0.97 (t, J = 7.2
Hz, 3H); LCMS: 95.7%, MS (ESI): m/z 531.1 [M + H]+. 39 ##STR00247##
white solid; .sup.1H-NMR (CDCl3, 400 MHz): .delta. 9.27 (s, 1H),
8.48- 8.54 (m, 2H), 8.29 (d, J = 2.4 Hz, 1H), 7.62 (m, 1H), 7.56
(s, 1H), 7.48 (s, 2H), 6.48 (d, J = 5.2 Hz, 1H), 4.56 (q, J = 8.4
Hz, 2H), 4.41 (t, J = 6.4 Hz, 2H), 4.08 (s, 6H), 3.04 (t, J = 6.4
Hz, 2H); LCMS: 94.1%, MS (ESI): m/z 543.2 [M + H]+. 40 ##STR00248##
yellow solid; .sup.1H-NMR (CDCl3, 400 MHz): .delta. 9.46 (s, 1H),
8.64 (d, J = 9.2 Hz, 1H), 8.53 (t, J = 6.0 Hz, 1H), 8.33 (d, J =
2.8 Hz, 1H), 8.20 (s, 1H), 7.63-7.71 (m, 2H), 7.37 (s, 1H), 6.75
(d, J = 6.4 Hz, 1H), 4.63 (q, J = 7.2 Hz, 1H), 4.55 (q, J = 8.4 Hz,
2H), 4.19 (s, 3H), 4.13 (s, 3H), 2.16-2.28 (m, 2H), 2.01-2.12 (m,
2H), 1.87-1.98 (m, 2H), 1.73-1.80 (m, 2H); LCMS: 96.4%, MS (ESI):
m/z 558.2 [M + H]+. 41 ##STR00249## yellow solid; .sup.1H-NMR
(CDCl3, 400 MHz): .delta. 9.51 (s, 1H), 8.65 (d, J = 9.2 Hz, 1H),
8.53-8.56 (m, 1H), 8.34 (d, J = 2.8 Hz, 1H), 8.22 (s, 1H),
7.67-7.70 (m, 2H), 7.47 (s, 1H), 6.76 (d, J = 6.4 Hz, 1H), 4.58 (q,
J = 8.0 Hz, 2H), 4.21 (s, 3H), 4.14 (s, 3H), 4.01 (d, J = 7.2 Hz,
2H), 1.28-1.35 (m, 1H), 0.73-0.78 (m, 2H), 0.44- 0.48 (m, 2H);
LCMS: 98.0%, MS (ESI): m/z 544.3 [M + H]+. 42 ##STR00250##
off-white solid; .sup.1H-NMR (DMSO-d6, 400 MHz): .delta. 9.59 (s,
1H), 8.83 (d, J = 6.4 Hz, 1H), 8.51 (d, J = 2.8 Hz, 1H), 8.39 (d, J
= 8.8 Hz, 1H), 8.00 (dd, J = 2.4, 8.8 Hz, 1H), 7.96 (s, 1H), 7.77
(s, 1H), 7.61 (s, 1H), 7.01 (d, J = 6.4 Hz, 1H), 4.78 (q, J = 8.8
Hz, 2H), 4.17 (d, J = 7.6 Hz, 2H), 4.04 (s, 3H), 4.03 (s, 3H),
2.76- 2.83 (m, 1H), 2.00-2.02 (m, 2H), 1.79-1.87 (m, 4H); LCMS:
98.6%, MS (ESI): m/z 558.3 [M + H]+. 43 ##STR00251## yellow solid;
.sup.1H-NMR (CDCl3, 400 MHz): .delta. 9.44 (s, 1H), 8.63 (d, J =
8.8 Hz 1H), 8.54 (d, J = 6.4 Hz, 1H), 8.34 (d, J = 2.8 Hz, 1H),
8.20 (s, 1H), 7.66 -7.69 (m, 2H), 7.31 (s, 1H), 6.75 (d, J = 6.4
Hz, 1H), 4.56 (q, J = 8.4 Hz, 2H), 4.20 (s, 3H), 4.14 (s, 3H), 3.94
(d, J = 7.2 Hz, 2H), 1.93-1.94 (m, 1H), 1.75-1.80 (m, 3H),
1.59-1.66 (m, 2H), 1.21-1.30 (m, 3H), 0.99-1.02 (m, 2H); LCMS:
96.3%, MS (ESI): m/z 586.3 [M + H]+. 44 ##STR00252## yellow solid;
.sup.1H-NMR (CDCl3, 400 MHz): .delta. 9.68 (s, 1H), 8.67 (d, J =
9.2 Hz, 1H), 8.55 (s, 1H), 8.33 (s, 1H), 8.19 (s, 1H), 7.72 (d, J =
7.2 Hz, 1H), 7.64 (s, 1H), 7.36 (s, 1H), 6.75 (d, J = 6.0 Hz, 1H),
4.55 (q, J = 8.4 Hz, 2H), 4.18 (s, 3H), 4.12 (s, 3H), 4.10 (s, 2H),
3.92-3.99 (s, 1H), 3.75-3.82 (m, 2H), 3.56-3.63 (m, 1H), 2.89-2.94
(m, 1H), 2.08-2.12 (m, 1H), 1.66-1.70 (m, 1H); LCMS: 96.2%, MS
(ESI): m/z 574.2 [M + H]+. 45 ##STR00253## white solid; .sup.1H-NMR
(CDCl3, 400 MHz): .delta. 8.88 (d, J = 2.4 Hz, 1H), 8.68 (t, J =
8.8 Hz, 1H), 8.43 (d, J = 6.4 Hz, 1H), 8.08 (d, J = 3.6 Hz, 1H),
7.55 (s, 1H), 7.24 (s, 1H), 7.01-7.03 (m, 2H), 6.68-6.69 (m, 1H),
4.42-4.48 (q, J = 8.0 Hz, 2H), 4.10 (s, 3H), 4.04 (s, 3H), 3.88 (s,
3H); LCMS: 97.1%, MS (ESI): m/z 521.2 [M + H]+. 46 ##STR00254##
white solid; .sup.1H-NMR (CDCl3, 400 MHz): .delta. 8.96 (d, J = 2.4
Hz, 1H), 8.75-8.80 (m, 1H), 8.51 (d, J = 8.0 Hz, 1H), 8.17 (s, 1H),
7.64 (s, 1H), 7.49 (s, 1H), 7.09-7.14 (m, 2H), 6.79 (d, J = 4.0 Hz,
1H), 4.52 (q, J = 8.00 Hz, 2H), 4.31-4.34 (m, 2H) 4.20 (s, 3H)
4.12-4.13 (m, 5H), 2.41 (s, 1H); LCMS: 97.4%, MS (ESI): m/z 551.2
[M + H]+. 47 ##STR00255## yellow solid; .sup.1H-NMR (CDCl3, 400
MHz): .delta. 9.00 (br s, 1H), 8.75 (t, J = 8.0 Hz, 1H), 8.51 (s,
1H), 8.17 (s, 1H), 7.63 (s, 1H), 7.36 (s, 1H), 7.10 (d, J = 8.8 Hz,
2H), 6.78 (s, 1H), 4.66 (q, J = 7.2 Hz, 1H), 4.54 (q, J = 8.4 Hz,
2H), 4.18 (s, 3H), 4.12 (s, 3H), 2.16-2.25 (m, 2H), 1.99-2.10 (m,
2H), 1.85-1.97 (m, 2H), 1.72-1.82 (m, 2H); LCMS: 96.2%, MS (ESI):
m/z 575.2 [M + H]+. 48 ##STR00256## white solid; .sup.1H-NMR
(CDCl3, 400 MHz): .delta. 8.91 (d, J = 2.4 Hz, 1H), 8.68 (t, J =
8.8 Hz, 1H), 8.44 (d, J = 4.8 Hz, 1H), 8.11 (s, 1H), 7.55 (s, 1H),
7.23 (s, 1H), 7.03 (s, 1H), 7.01 (s, 1H), 6.69 (d, J = 6.8 Hz, 1H),
4.46 (q, J = 8.4 Hz, 2H), 4.11 (s, 3H), 4.04 (s, 3H), 3.85 (d, J =
7.2 Hz, 2H), 2.14-2.21 (m, 1H), 0.90 (s, 3H), 0.88 (s, 3H); LCMS:
100%, MS (ESI): m/z 563.2 [M + H]+. 49 ##STR00257## yellow solid;
.sup.1H-NMR (CDCl3, 400 MHz): .delta. 8.97 (d, J = 2.4 Hz, 1H),
8.77 (t, J = 8.8 Hz, 1H), 8.52-8.53 (m, 1H), 8.18 (s, 1H), 7.63 (s,
1H), 7.44 (s, 1H), 7.09-7.11 (m, 2H), 6.77 (d, J = 3.6 Hz, 1H),
4.52 (q, J = 8.0 Hz, 2H), 4.32 (t, J = 4.8 Hz, 2H), 4.18 (s, 3H),
4.12 (s, 3H), 3.78 (t, J = 4.8 Hz, 2H), 3.38 (s, 3H); LCMS: 100%,
MS (ESI): m/z 565.3 [M + H]+. 50 ##STR00258## white solid;
.sup.1H-NMR (CDCl3, 400 MHz): .delta. 8.62 (s, 1H), 8.43 (d, J =
5.2 Hz, 1H), 7.82 (dd, J = 2.4, 12.0 Hz, 1H), 7.53 (s, 1H), 7.38
(s, 2H), 7.24-7.30 (m, 2H), 6.37 (d, J = 5.2 Hz, 1H), 4.48 (q, J =
8.4 Hz, 2H), 4.00 (s, 3H), 3.99 (s, 3H), 3.86 (s, 3H); LCMS: 98.2%,
MS (ESI): m/z 521.2 [M + H]+. 51 ##STR00259## gray solid;
.sup.1H-NMR (CDCl3, 400 MHz): .delta. 8.69 (s, 1H), 8.43 (d, J =
8.0 Hz, 1H), 8.06 (s, 1H), 7.90 (dd, J = 12.10, 2.4 Hz, 1H), 7.59
(s, 1H), 7.38-7.40 (m, 2H), 7.24 (t, J = 8.0 Hz, 1H), 6.65 (d, J =
8.0 Hz, 1H), 4.45 (q, J = 8.0 Hz, 2H), 4.20-4.23 (m, 2H), 4.10 (s,
3H), 4.00-4.02 (m, 5H); LCMS: 95.6%, MS (ESI): m/z 551.2 [M + H]+.
52 ##STR00260## yellow solid; .sup.1H-NMR (CDCl3, 400 MHz): .delta.
8.79 (s, 1H), 8.52 (t, J = 6.0 Hz, 1H), 8.18 (s, 1H), 7.99 (dd, J =
2.4, 12.4 Hz, 1H), 7.67 (s, 1H), 7.48 (d, J = 8.8 Hz, 1H), 7.37 (s,
1H), 7.32 (t, J = 8.4 Hz, 1H), 6.74 (d, J = 6.0 Hz, 1H), 4.64 (q, J
= 7.2 Hz, 1H), 4.55 (q, J = 8.4 Hz, 2H), 4.18 (s, 3H), 4.13 (s,
3H), 2.16-2.28 (m, 2H), 1.99-2.09 (m, 2H), 1.86-1.96 (m, 2H),
1.74-1.83 (m, 2H); LCMS: 99.2%, MS (ESI): m/z 575.2 [M + H]+. 53
##STR00261## yellow solid; .sup.1H-NMR (CDCl3, 400 MHz): .delta.
8.71 (s, 1H), 8.40- 8.48 (m, 1H), 8.10 (s, 1H), 7.92 (dd, J = 2.0,
12.0 Hz, 1H), 7.59 (s, 1H), 7.41 (d, J = 8.0 Hz, 1H), 7.22-7.26 (m,
2H), 6.67 (d, J = 5.2 Hz, 1H), 4.48 (q, J = 8.4 Hz, 2H), 4.11 (s,
3H), 4.05 (s, 3H), 3.84 (d, J = 7.6 Hz, 2H), 2.12-2.22 (m, 1H),
0.90 (s, 3H), 0.88 (s, 3H); LCMS: 98.3%, MS (ESI): m/z 563.2 [M +
H]+. 54 ##STR00262## white solid; .sup.1H-NMR (CDCl3, 400 MHz):
.delta. 8.77 (s, 1H), 8.52 (t, J = 3.2 Hz, 1H), 8.19 (s, 1H), 7.99
(dd, J = 2.4, 12.0 Hz, 1H), 7.67 (s, 1H), 7.42-7.49 (m, 2H), 7.32
(t, J = 8.4 Hz, 1H), 6.74 (d, J = 6.0 Hz, 1H), 4.54 (q, J = 8.4 Hz,
2H), 4.30 (t, J = 4.8 Hz, 2H), 4.19 (s, 3H), 4.13 (s, 3H), 3.77 (t,
J = 4.8 Hz, 2H), 3.39 (s, 3H); LCMS: 97.6%, MS (ESI): m/z 565.2 [M
+ H]+. 55 ##STR00263## yellow solid; .sup.1H-NMR (CDCl3, 400 MHz):
.delta. 8.79 (s, 1H), 8.54 (d, J = 6.0 Hz, 1H), 8.18 (s, 1H), 8.01
(d, J = 2.0, 12.0 Hz, 1H), 7.68 (s, 1H), 7.47-7.50 (m, 2H), 7.34
(t, J = 8.4 Hz, 1H), 6.78 (d, J = 6.0 Hz, 1H), 4.99-5.02 (m, 1H),
4.55 (q, J = 8.4 Hz, 2H), 4.16-4.22 (m, 5H), 4.14 (s, 1H),
4.03-4.07 (m, 1H), 3.94-4.01 (m, 1H), 2.53-2.62 (m, 1H), 2.30-2.38
(m, 1H); LCMS: 94.1%, MS (ESI): m/z 577.2 [M + H]+. 56 ##STR00264##
yellow solid; .sup.1H-NMR (CDCl3, 400 MHz): .delta. 8.70 (s, 1H),
8.47 (t, J = 8.0 Hz, 1H), 8.16 (s, 1H), 7.78 (s, 1H), 7.67-7.70 (m,
2H), 7.49 (s, 1H), 7.17 (d, J = 8.0 Hz, 1H), 6.62 (d, J = 4.0 Hz,
1H), 4.55 (q, J = 8.4 Hz, 2H), 4.29-4.32 (m, 2H), 4.19 (s, 3H),
4.14- 4.19 (m, 3H), 4.09-4.12 (m, 2H), 2.21 (s, 3H); LCMS: 99.1%,
MS (ESI): m/z 547.2 [M + H]+. 57 ##STR00265## yellow solid;
.sup.1H-NMR (CDCl3, 400 MHz): .delta. 8.71 (s, 1H), 8.47 (t, J =
6.4 Hz, 1H), 8.16 (s, 1H), 7.79 (d, J = 2.0 Hz, 1H), 7.65- 7.68 (m,
2H), 7.37 (s, 1H) 7.15 (d, J = 8.8 Hz, 1H), 6.62 (d, J = 6.4 Hz,
1H), 4.64 (q, J =6.8 Hz, 1H), 4.55 (q, J = 8.4 Hz, 2H), 4.18 (s,
3H), 4.13 (s, 3H), 2.20-2.23 (m, 5H), 2.04-2.05 (m, 2H), 1.91-1.92
(m, 2H), 1.76-1.79 (m, 2H); LCMS: 98.4%, MS (ESI): m/z 571.2 [M +
H]+. 58 ##STR00266## white solid; .sup.1H-NMR (CDCl3, 400 MHz):
.delta. 8.63 (s, 1H), 8.36- 8.42 (m, 1H), 8.10 (s, 1H), 7.71 (d, J
= 2.4 Hz, 1H), 7.58-7.60 (m, 2H), 7.24 (s, 1H), 7.07 (d, J = 8.8
Hz, 1H), 6.54 (d, J = 6.4 Hz, 1H), 4.49 (q, J = 8.4 Hz, 2H), 4.10
(s, 3H), 4.05 (s, 3H), 3.84 (d, J = 7.2 Hz, 2H), 2.12-2.22 (m, 4H),
0.90 (s, 3H), 0.88 (s, 3H); LCMS: 98.2%, MS (ESI): m/z 559.3 [M +
H]+. 59 ##STR00267## yellow solid; .sup.1H-NMR (CDCl3, 400 MHz):
.delta. 8.75 (s, 1H), 8.40- 8.50 (m, 1H), 8.13 (s, 1H), 7.88 (d, J
= 8.4 Hz, 2H), 7.66 (s, 1H), 7.48 (s, 1H), 7.25 (s, 1H), 6.73 (d, J
= 4.4 Hz, 1H), 4.50 (q, J = 8.4 Hz, 2H), 4.29-4.30 (m, 2H), 4.18
(s, 3H), 4.10-4.12 (m, 5H), 1.26 (s, 1H); LCMS: 98.2%, MS (ESI):
m/z 533.2 [M + H]+. 60 ##STR00268## yellow solid; .sup.1H-NMR
(CDCl3, 400 MHz): .delta. 8.76 (s, 1H), 8.48 (t, J = 6.4 Hz, 1H),
8.16 (s, 1H), 7.89 (d, J = 8.8 Hz, 2H), 7.66 (s, 1H), 7.37 (s, 1H),
7.22-7.30 (m, 2 H), 6.74 (d, J = 6.4 Hz, 1H), 4.65 (q, J = 7.2 Hz,
1H), 4.56 (q, J = 8.4 Hz, 2H), 4.18 (s, 3H), 4.12 (s, 3H), 2.15
2.27 (m, 2H), 1.98-2.10 (m, 2H), 1.84-1.97 (m, 2H), 1.70-1.84 (m,
2H); LCMS: 95.0%, MS (ESI): m/z 557.2 [M + H]+. 61 ##STR00269##
white solid; .sup.1H-NMR (CDCl3, 400 MHz): .delta. 8.75 (s, 1H),
8.48- 8.49 (m, 1H), 8.17 (s, 1H), 7.88 (d, J = 8.8 Hz, 2H), 7.66
(s, 1H), 7.45 (s, 1H), 7.24 (d, J = 8.8 Hz, 2H), 6.74 (d, J = 6.4
Hz, 1H), 4.55 (q, J = 8.0 Hz, 2H), 4.30 (t, J = 5.2 Hz, 2H), 4.18
(s, 3H), 4.12 (s, 3H), 3.77 (t, J = 5.2 Hz, 2H), 3.39 (s, 3H);
LCMS: 98.4%, MS (ESI): m/z 547.2 [M + H]+. 62 ##STR00270## yellow
solid; .sup.1H-NMR (CDCl3, 400 MHz): .delta. 8.67 (s, 1H), 8.38-
8.47 (m, 1H), 8.08 (s, 1H), 7.81 (d, J = 8.4 Hz, 2H), 7.58 (s, 1H),
7.38 (s, 1H), 7.18 (s, 1H), 7.16 (s, 1H), 6.64-6.70 (m, 1H), 4.88-
4.96 (m, 1H), 4.48 (q, J = 8.4 Hz, 2H), 4.07-4.13 (m, 5H), 4.04 (s,
1H), 3.95-3.99 (m, 1H), 3.85-3.91 (m, 1H), 2.43-2.53 (m, 1H),
2.24-2.28 (m, 1H); LCMS: 94.4%, MS (ESI): m/z 559.2 [M + H]+. 63
##STR00271## HCl salt, yellow solid; .sup.1H-NMR (MeOD, 400 MHz):
.delta. 8.77 (d, J = 6.8 Hz, 1H), 8.59 (d, J = 2.4 Hz, 1H), 8.49
(d, J = 9.2 Hz, 1H), 8.21 (dd, J = 9.2, 2.8 Hz, 1H), 7.92 (s, 1H),
7.86 (s, 1H), 7.54 (s, 1H), 7.15 (d, J = 6.8 Hz, 1H), 4.69 (q, J =
8.4 Hz, 2H), 4.40-4.56 (m, 2H), 4.15 (s, 3H), 4.11 (s, 3H),
3.88-3.98 (m, 1H), 1.38 (d, J = 6.4 Hz, 3H); LCMS: 99.2%, MS (ESI):
m/z 547.2 [M + H]+. 64 ##STR00272## yellow solid; .sup.1H-NMR
(MeOD, 400 MHz): .delta. 8.72 (d, J = 6.8 Hz, 1H), 8.40-8.59 (m,
2H), 8.02 (dd, J = 2.8, 9.2 Hz, 1H), 7.86 (s, 1H), 7.84 (s, 1H),
7.50 (s, 1H), 7.07 (d, J = 6.8 Hz, 1H), 4.63- 4.70 (m, 4H), 4.14
(s, 3H), 4.10 (s, 3H), 1.67 (t, J = 18.8 Hz, 3H); LCMS: 96.9%, MS
(ESI): m/z 568.3 [M + H]+. 65 ##STR00273## white solid; .sup.1H-NMR
(CDCl3, 400 MHz): .delta. 9.34 (s, 1H), 8.49- 8.54 (m, 2H),
8.26-8.29 (m, 1H), 7.61 (dd, J = 2.4, 8.8 Hz, 1H), 7.58 (s, 1H),
7.46 (s, 2H), 6.47 (d, J = 5.2 Hz, 1H), 4.55 (q, J = 8.4 Hz, 2H),
4.25 (t, J = 4.8 Hz, 2H), 4.08 (s, 6H), 3.88-3.93 (m, 1H), 3.70 (t,
J = 4.8 Hz, 2H), 2.16-2.20 (m, 2H), 1.83-1.90 (m, 2H), 1.68-1.72
(m, 1H), 1.49-1.54 (m, 1H); LCMS: 100%, MS (ESI): m/z 588.3 [M +
H]+. 66 ##STR00274## white solid; .sup.1H-NMR (MeOD, 400 MHz):
.delta. 8.43-8.46 (m, 2H), 8.33 (d, J = 2.4 Hz, 1H), 7.90 (s, 1H),
7.78 (dd, J = 2.4, 9.2 Hz, 1H), 7.63 (s, 1H), 7.36 (s, 1H), 6.60
(d, J = 5.2 Hz, 1H), 5.56 (q, J = 6.8 Hz, 1H), 5.05-5.10 (m, 4H),
4.65 (q, J = 8.4 Hz, 2H), 4.02 (s, 3H), 4.01 (s, 3H); LCMS: 97.0%,
MS (ESI): m/z 546.1 [M + H]+. 67 ##STR00275## white solid;
.sup.1H-NMR (MeOD, 400 MHz): .delta. 8.76 (d, J = 6.4 Hz, 1H), 8.56
(d, J = 2.4 Hz, 1H), 8.36 (d, J = 9.2 Hz, 1H), 8.21 (dd, J = 2.8,
9.2 Hz, 1H), 7.86 (s, 1H), 7.80 (s, 1H), 7.52 (s, 1H), 6.15 (d, J =
6.8 Hz, 1H), 4.64 (q, J = 8.4 Hz, 2H), 4.31 (t, J = 4.8 Hz, 2H),
4.14 (s, 3H), 4.10 (s, 3H), 3.97 (t, J = 4.2 Hz, 2H); LCMS: 99.4%,
MS (ESI): m/z 534.2 [M + H]+. 68 ##STR00276## HCl salt, white
solid; .sup.1H-NMR (DMSO-d6, 400 MHz): .delta. 9.68 (s, 1H), 8.85
(d, J = 6.4 Hz, 1H), 8.53 (d, J = 2.8 Hz, 1H), 8.39 (d, J = 9.2 Hz,
1H), 8.14 (s, 2H), 8.01-8.04 (m, 2H), 7.77 (d, J = 10.8 Hz, 2H),
7.04 (d, J = 6.4 Hz, 1H), 4.78 (q, J = 8.8 Hz, 2H), 4.27 (t, J =
6.4 Hz, 3H), 4.04 (s, 6H), 2.80-2.82 (m, 2H), 2.13-2.17 (m, 2H);
LCMS: 98.4%, MS (ESI): m/z 547.1 [M + H]+. 69 ##STR00277## yellow
solid; .sup.1H-NMR (CDCl3, 400 MHz): .delta. 9.44 (s, 1H), 8.62 (d,
J = 9.2 Hz, 1H), 8.53 (d, J = 6.4 Hz, 1H), 8.33 (d, J = 2.4 Hz,
1H), 8.19 (s, 1H), 7.64-7.67 (m, 2H), 7.36 (s, 1H), 6.75 (d, J =
6.4 Hz, 1H), 4.54 (q, J = 8.4 Hz, 2H), 4.19 (s, 3H), 4.13 (s, 3H),
4.05-4.09 (m, 1H), 2.17-2.21 (m, 2H), 1.94-1.98 (m, 2H), 1.71- 1.81
(m, 3H), 1.41-1.52 (m, 2H), 1.25-1.35 (m, 1H); LCMS: 96.7%, MS
(ESI): m/z 572.2 [M + H]+. 70 ##STR00278## HCl salt, yellow solid;
.sup.1H-NMR (MeOD, 400 MHz): .delta. 8.77 (d, J = 6.8 Hz, 1H),
8.59-8.64 (m, 2H), 8.26 (dd, J = 9.2, 2.8 Hz, 1H), 7.96 (s, 1H),
7.86 (s, 1H), 7.54 (s, 1H), 7.17 (d, J = 6.4 Hz, 1H), 5.34 (br t, J
= 6.4 Hz, 1H), 4.68 (q, J = 8.4 Hz, 2H), 4.14 (s, 3H), 4.10 (s,
3H), 3.98 (d, J = 12.8 Hz, 1H), 3.66-3.79 (m, 2H), 3.48- 3.61 (m,
1H), 2.55-2.69 (m, 1H), 2.33-2.45 (m, 1H); LCMS: 97.1%, MS (ESI):
m/z 559.2 [M + H]+. 71 ##STR00279## off-white solid; .sup.1H-NMR
(CDCl3, 400 MHz): .delta. 9.44 (s, 1H), 8.62 (d, J = 9.2 Hz, 1H),
8.53 (s, 1H), 8.33 (d, J = 2.8 Hz, 1H), 8.20 (s, 1H), 7.66-7.68 (m,
2H), 7.34 (s, 1H), 6.74 (d, J = 6.4 Hz, 1H), 4.55 (q, J = 8.4 Hz,
2H), 4.19 (s, 3H), 4.13 (s, 3H), 4.04 (d, J
= 7.2 Hz, 2H), 2.42-2.50 (m, 1H), 1.61-1.77 (m, 6H), 1.26- 1.29 (m,
2H).; LCMS: 97.5%, MS (ESI): m/z 572.2 [M + H]+. 72 ##STR00280##
HCl salt, white solid; .sup.1H-NMR (MeOD, 400 MHz): .delta. 8.78
(d, J = 6.8 Hz, 1H), 8.60 (d, J = 2.8 Hz, 1H), 8.45 (d, J = 9.2 Hz,
1H), 8.23 (dd, J = 2.8, 9.2 Hz, 1H), 7.92 (s, 1H), 7.87 (s, 1H),
7.56 (s, 1H), 7.18 (d, J = 6.8 Hz, 1H), 4.69 (q, J = 8.4 Hz, 2H),
4.38-4.40 (m, 2H), 4.16 (s, 3H), 4.12 (s, 3H), 3.47-3.52 (m, 2H),
3.35- 3.38 (m, 1H), 3.19-3.24 (m, 1H), 3.06-3.08 (m, 1H), 2.22-2.26
(m, 1H), 1.87-1.93 (m, 1H); LCMS: 98.2%, MS (ESI): m/z 573.2 [M +
H]+. 73 ##STR00281## HCl salt, white solid; .sup.1H-NMR (DMSO-d6,
400 MHz): .delta. 9.73 (s, 1H), 9.41 (s, 1H), 8.84 (d, J = 6.8 Hz,
1H), 8.53 (d, J = 2.8 Hz, 1H), 8.39 (d, J = 9.2 Hz, 1H), 8.00-8.04
(m, 2H), 7.82 (s, 1H), 7.78 (s, 1H), 7.03 (d, J = 6.8 Hz, 1H), 4.79
(q, J = 8.8 Hz, 2H), 4.47 (d, J = 7.2 Hz, 1H), 3.99-4.05 (m, 8H),
3.82-3.90 (m, 2H), 3.23-3.31 (m, 1H); LCMS: 99.0%, MS (ESI): m/z
559.2 [M + H]+. 74 ##STR00282## HCl salt, white solid; .sup.1H-NMR
(MeOD, 400 MHz): .delta. 8.71 (d, J = 6.8 Hz, 1H), 8.46 (t, J = 8.8
Hz, 1H), 7.95 (s, 1H), 7.83 (s, 1H), 7.51 (s, 1H), 7.45 (dd, J =
10.8, 2.4 Hz, 1H), 7.31 (br d, J = 8.8 Hz, 1H), 7.04 (d, J = 6.8
Hz, 1H), 5.31 (br t, J = 6.4 Hz, 1H), 4.72 (q, J = 8.4 Hz, 2H),
4.13 (s, 3H), 4.10 (s, 3H), 3.90 (d, J = 12.8 Hz, 1H), 3.53-3.76
(m, 3H), 2.66 (s, 1H), 2.57-2.65 (m, 1H), 2.34-2.43 (m, 1H); LCMS:
97.7%, MS (ESI): m/z 576.2 [M + H]+. 75 ##STR00283## HCl salt,
off-white solid; .sup.1H-NMR (D.sub.2O, 400 MHz): .delta. 8.43 (d,
J = 6.8 Hz, 1H), 7.86 (t, J = 8.8 Hz, 1H), 7.65 (s, 1H), 7.55 (s,
1H), 7.23 (s, 1H), 7.11 (d, J = 10.4 Hz, 1H), 7.05 (d, J = 9.2 Hz,
1H), 6.87 (d, J = 6.8 Hz, 1H), 4.46 (q, J = 8.0 Hz, 2H), 4.38 (s,
2H), 3.90 (s, 3H), 3.85 (s, 3H), 3.42 (t, J = 5.2 Hz, 2H); LCMS:
99.0%, MS (ESI): m/z 550.2 [M + H]+. 76 ##STR00284## yellow solid;
.sup.1H-NMR (MeOD, 400 MHz): .delta. 8.60 (d, J = 6.8 Hz, 1H), 8.28
(t, J = 8.8 Hz, 1H), 7.73 (s, 1H), 7.66 (s, 1H), 7.39 (s, 1H), 7.31
(dd, J = 1.6,10.8 Hz, 1H), 7.17 (d, J = 8.4 Hz, 1H), 6.93 (d, J =
6.8 Hz, 1H), 4.56 (q, J = 8.4 Hz, 2H), 4.07-4.12 (m, 2H), 4.03 (s,
3H), 3.99 (s, 3H), 3.94-3.98 (m, 1H), 1.12 (d, J = 6.0 Hz, 3H);
LCMS: 98.6%, MS (ESI): m/z 565.2 [M + H]+. 77 ##STR00285## yellow
solid; .sup.1H-NMR (CDCl3, 400 MHz): .delta. 8.95 (s, 1H), 8.75 (t,
J = 8.8 Hz, 1H), 8.51 (s, 1H), 8.16 (s, 1H), 7.62 (s, 1H), 7.43 (s,
1H), 7.09-7.11 (m, 2H), 6.77 (s, 1H), 4.98-5.06 (m, 1H), 4.52 (q, J
= 8.0 Hz, 2H), 4.07-4.21 (m, 8H), 4.03-4.05 (m, 1H), 3.94- 3.96 (m,
1H), 2.51-2.60 (m, 1H), 2.31-2.35 (m, 1H); LCMS: 97.7%, MS (ESI):
m/z 577.2 [M + H]+. 78 ##STR00286## HCl salt, white solid;
.sup.1H-NMR (MeOD, 400 MHz): .delta. 8.72 (d, J = 6.4 Hz, 1H),
8.11-8.14 (m, 1H), 7.89 (s, 1H), 7.86 (s, 1H), 7.76- 7.74 (m, 1H),
7.47-7.56 (m, 2H), 7.01 (d, J = 6.8 Hz, 1H), 5.29 (s, 1H),
4.64-4.87 (m, 2H), 4.14 (s, 3H), 4.11 (s, 3H), 3.93-3.97 (m, 1 H),
3.68-3.71 (m, 2H), 3.54-3.57 (m, 1H), 2.59-2.64 (m, 1H), 2.40-2.41
(m, 1H); LCMS: 95.7%, MS (ESI): m/z 576.2 [M + H]+. 79 ##STR00287##
HCl salt, white solid; .sup.1H-NMR (DMSO-d6, 400 MHz): .delta.
10.71 (s, 1H), 8.83 (d, J = 6.4 Hz, 1H), 8.57 (s, 3H, NH2.HCl),
8.21 (dd, J = 2.4, 13.2 Hz, 1H), 7.99 (s, 1H), 7.92 (d, J = 9.6 Hz,
1H), 7.81 (s, 1H), 7.77 (s, 1H), 7.62 (t, J = 9.2 Hz, 1H), 6.98 (d,
J = 6.4 Hz, 1H), 4.70 (q, J = 8.8 Hz, 2H), 4.47 (t, J = 5.6 Hz,
2H), 4.05 (s, 3H), 4.04 (s, 3H), 3.35 (d, J = 5.6 Hz, 2H); LCMS:
98.8%, MS (ESI): m/z 550.2 [M + H]+. 80 ##STR00288## yellow solid;
.sup.1H-NMR (MeOD, 400 MHz): .delta. 8.71 (d, J = 6.8 Hz, 1H), 8.09
(dd, J = 2.4, 12.8 Hz, 1H), 7.87 (s, 1H), 7.73 (s, 1H), 7.65-7.68
(m, 1H), 7.31 (t, J = 8.8 Hz, 1H), 7.48 (s, 1H), 7.02 (d, J = 6.4
Hz, 1H), 4.63 (q, J = 8.4 Hz, 2H), 4.20-4.24 (m, 2H), 4.15 (s, 3H),
4.12 (s, 3H), 4.08 (q, J = 8.4 Hz, 1H), 1.23 (d, J = 6.0 Hz, 3H);
LCMS: 98.2%, MS (ESI): m/z 565.2 [M + H]+. 81 ##STR00289## HCl
salt, white solid; .sup.1H-NMR (MeOD, 400 MHz): .delta. 8.68 (d, J
= 6.8 Hz, 1H), 7.89-7.92 (m, 3H), 7.85 (dd, J = 1.8, 8.8 Hz, 1H),
7.54 (s, 1H), 7.31 (d, J = 8.8 Hz, 1H), 6.85 (d, J = 6.8 Hz, 1H),
5.32 (t, J = 6.0 Hz, 1H), 4.68 (q, J = 8.4 Hz, 2H), 4.15 (s, 3H),
4.13 (s, 3H), 3.97 ( d, J = 12.8 Hz, 1H), 3.71-3.77 (m, 2H),
3.53-3.63 (m, 1H), 2.61-2.66 (m, 1H), 2.41-2.43 (m, 1H), 2.25 (s,
3H); LCMS: 98.5%, MS (ESI): m/z 572.2 [M + H]+. 82 ##STR00290## HCl
salt, white solid; .sup.1H-NMR (DMSO-d6, 400 MHz): .delta. 10.33
(s, 1H), 8.78 (d, J = 6.8 Hz, 1H), 8.54 (s, 3H, NH2.HCl), 7.99 (d,
J = 2.0 Hz, 1H), 7.98 (s, 1H), 7.87 (dd, J = 2.4, 8.8 Hz, 1H), 7.80
(s, 1H), 7.79 (s, 1H), 7.33 (d, J = 8.8 Hz, 1H), 6.74 (d, J = 6.4
Hz, 1H), 4.71 (q, J = 8.8 Hz, 2H), 4.46 (t, J = 5.6 Hz, 2H), 4.05
(s, 3H), 4.04 (s, 3H), 3.49 (q, J = 5.2 Hz, 2H), 2.15 (s, 3H);
LCMS: 98.9%, MS (ESI): m/z 546.2 [M + H]+. 83 ##STR00291## yellow
solid; .sup.1H-NMR (CDCl3, 400 MHz): .delta. 8.65 (s, 1H), 8.39 (s,
1H), 8.04 (s, 1H), 7.67 (s, 1H), 7.61-7.62 (m, 2H), 7.44 (s, 1H),
7.07 (d, J = 8.8 Hz, 1H), 6.55 ( s, 1H), 4.43 (q, J = 8.4 Hz, 2H),
4.24 (br.s., 1H), 4.09-4.14 (m, 4H), 4.05 (s, 3H), 3.98-3.99 (m,
1H), 2.64 (s, 1H), 2.11 (s, 3H), 1.22 (d, J = 6.0 Hz, 3H); LCMS:
97.1%, MS (ESI): m/z 561.2 [M + H]+. 84 ##STR00292## white solid;
.sup.1H-NMR (CDCl3, 400 MHz): .delta. 8.62 (s, 1H), 8.40 (s, 1H),
8.09 (s, 1H), 7.71 (d, J = 1.6 Hz, 1H), 7.58-7.60 (m, 2H), 7.38 (s,
1H), 7.07 (d, J = 8.4 Hz, 1H), 6.54 (d, J = 6.0 Hz, 1H), 4.47 (q, J
= 8.0 Hz, 2H), 4.22 (t, J = 5.2 Hz, 2H), 4.10 (s, 3H), 4.05 (S,
3H), 3.69 (t, J = 4.8 Hz, 2H), 3.31 (s, 3H), 2.12 (s, 3H); LCMS:
98.9%, MS (ESI): m/z 561.2 [M + H]+. 85 ##STR00293## yellow solid;
.sup.1H-NMR (CDCl3, 400 MHz): .delta. 8.62 (s, 1H), 8.41 (t, J =
6.0 Hz, 1H), 8.08 (s, 1H), 7.70 (d, J = 2.4 Hz, 1H), 7.61 (s, 1H),
7.58 (s, 1H), 7.39 (s, 1H), 7.07 (d, J = 8.8 Hz, 1H), 6.54 (d, J =
6.0 Hz, 1H), 4.90-4.93 (m, 1H), 4.48 (q, J = 8.4 Hz, 2H), 4.12-4.13
(m, 1H), 4.09 (s, 3H), 4.07 (s, 1H), 4.05 (s, 3H), 3.95-3.99 (m,
1H), 3.85-3.91 (m, 1H), 2.45-2.52 (m, 1H), 2.25- 2.28 (m, 1H), 2.10
(s, 3H); LCMS: 98.2%, MS (ESI): m/z 573.3 [M + H]+. 86 ##STR00294##
HCl salt, yellow solid; .sup.1H-NMR (MeOD, 400 MHz): .delta. 8.68
(d, J = 6.8 Hz, 1H), 8.00 (m, J = 9.2 Hz, 2H), 7.89 (s, 1H), 7.85
(s, 1H), 7.48 (s, 1H), 7.40 (m, J = 8.8 Hz, 2H), 6.95 (d, J = 6.8
Hz, 1H), 5.29 (t, J = 6.4 Hz, 1H), 4.67 (q, J = 8.4 Hz, 2H), 4.13
(s, 3H), 4.10 (s, 3H), 3.94 (d, J = 12.8 Hz, 1H), 3.64-3.75 (m,
2H), 3.48-3.59 (m, 1H), 2.56-2.67 (m, 1H), 2.34-2.43 (m, 1H); LCMS:
98.1%, MS (ESI): m/z 558.2 [M + H]+. 87 ##STR00295## white solid;
.sup.1H-NMR (CDCl3, 400 MHz): .delta. 8.78 (s, 1H), 8.50 (t, J =
6.0 Hz, 1H), 8.18 (s, 1H), 7.89-7.91 (m, 1H), 7.67 (s, 1H), 7.33
(s, 1H), 7.24-7.27 (m, 2H), 6.75 (d, J = 6.8 Hz, 1H), 4.58 (q, J =
8.4 Hz, 2H), 4.19 (s, 3H), 4.13 (s, 3H), 3.94 (s, 1H), 3.92 (s,
1H), 2.19-2.31 (m, 1H), 0.99 (s, 3H), 0.97 (s, 3H); LCMS: 99.6%, MS
(ESI): m/z 545.3 [M + H]+. 88 ##STR00296## HCl salt, white solid;
.sup.1H-NMR (DMSO-d6, 400 MHz): .delta. 10.41 (s, 1H), 8.79 (d, J =
6.4 Hz, 1H), 8.46 (s, 3H, NH2.HCl), 8.06 (d, J = 8.8 Hz, 2H), 7.98
(s, 1H), 7.74 (d, J = 8.8 Hz, 2H), 7.41 (d, J = 9.2 Hz, 2H), 6.85
(d, J = 6.4 Hz, 1H), 4.71 (q, J = 9.2 Hz, 2H), 4.45 (t, J = 5.6 Hz,
2H), 4.04 (s, 6H), 3.35 (d, J = 5.2 Hz, 2H); LCMS: 100%, MS (ESI):
m/z 532.2 [M + H]+. 89 ##STR00297## yellow solid; .sup.1H-NMR
(MeOD, 400 MHz): .delta. 8.68 (d, J = 6.8 Hz, 1H), 7.95 (d, J = 8.8
Hz, 2H), 7.86 (s, 1H), 7.73 (s, 1H), 7.50 (s, 1H), 7.39 ( d, J =
8.8 Hz, 2H), 6.96 (d, J = 6.8 Hz, 1H), 4.63 (q, J = 8.4 Hz, 2H),
4.19-4.26 (m, 2H), 4.14 (s, 3H), 4.09 (s, 3H), 4.03-4.08 (m, 1H),
1.23 (d, J = 6.0 Hz, 3H); LCMS: 97.6%, MS (ESI): m/z 547.2 [M +
H]+. 90 ##STR00298## white solid; .sup.1H-NMR (MeOD, 400 MHz):
.delta. 8.43-8.46 (m, 2H), 8.32 (d, J = 2.8 Hz, 1H), 7.83 (s, 1H),
7.78 (dd, J = 2.8, 9.2 Hz, 1H), 7.64 (s, 1H), 7.37 (s, 1H), 6.60
(d, J = 5.6 Hz, 2H), 4.65 (q, J = 8.8 Hz, 2H), 4.0-4.50 (m, 1H),
4.07-4.10 (m, 2H), 4.02 (s, 6H), 3.55-3.62 (m, 2H), 2.09-2.17 (m,
4H); LCMS: 97.1%, MS (ESI): m/z 574.2 [M + H]+. 91 ##STR00299##
yellow solid; .sup.1H-NMR (CDCl3, 400 MHz): .delta. 9.33 (s, 1H),
8.53 (t, J = 5.2 Hz, 1H), 8.49 (s, 1H), 8.28 (d, J = 2.8 Hz, 1H),
7.60 (dd, J = 2.8, 9.2 Hz, 1H), 7.57 (s, 1H), 7.56 (s, 1H), 7.45
(s, 1H), 6.47 (d, J = 5.6 Hz, 1H), 4.83-4.93 (m, 1H), 4.55 (q, J =
8.4 Hz, 2H), 4.07-4.08 (m, 6H), 2.95-3.00 (m, 2H), 2.80-2.82 (m,
1H), 2.45-2.48 (m, 2H), 2.43 (s, 3H), 2.15-2.17 (m, 1H), 1.62 (s,
2H); LCMS: 95.2%, MS (ESI): m/z 573.3 [M + H]+. 92 ##STR00300##
white solid; .sup.1H-NMR (MeOD, 400 MHz): .delta. 8.41-8.48 (m,
2H), 8.32 (d, J = 2.8 Hz, 1H), 7.74-7.81 (m, 2H), 7.64 (s, 1H),
7.37 (s, 1H), 6.59 (d, J = 5.6 Hz, 1H), 4.80-4.85 (m, 2H), 4.63 (q,
J = 8.4 Hz, 2H), 4.57 (q, J = 6.4 Hz, 2H), 4.50 (d, J = 7.2 Hz,
2H), 4.01 (s, 3H), 4.00 (s, 3H), 3.53-3.60 (m, 1H); LCMS: 99.4%, MS
(ESI): m/z 560.1 [M + H]+. 93 ##STR00301## yellow solid;
.sup.1H-NMR (MeOD, 400 MHz): .delta. 8.70 (d, J = 6.8 Hz, 1H), 8.54
(d, J = 9.2 Hz, 1H), 8.44 (d, J = 2.8 Hz, 1H), 7.91 (dd, J = 2.8,
8.8 Hz, 1H), 7.87 (s, 1H), 7.78 (s, 1H), 7.46 (s, 1H), 7.03 (d, J =
6.8 Hz, 1H), 4.65 (q, J = 8.4 Hz, 1H), 4.21 (t, J = 7.2 Hz, 2H),
4.13 (s, 3H), 4.10 (s, 3H), 1.94 (q, J = 7.2 Hz, 2H), 1.75-1.85 (m,
3H), 1.60-1.71 (m, 2H), 1.49-1.60 (m, 2H), 1.13- 1.25 (m, 2H);
LCMS: 100%, MS (ESI): m/z 586.3 [M + H]+. 94 ##STR00302## yellow
solid; .sup.1H-NMR (CDCl3, 400 MHz): .delta. 8.89 (s, 1H), 8.57-
8.62 (m, 1H), 8.53 (d, J = 5.2 Hz, 1H), 7.59 (s, 1H), 7.53 (s, 1H),
7.44 (s, 1H), 6.99-7.05 (m, 2H), 6.54 (d, J = 5.2 Hz, 1H), 4.89-
4.93 (m, 1H), 4.54 (q, J = 8.0 Hz, 2H), 4.07 (s, 3H), 4.06 (s, 3H),
2.96-3.03 (m, 2H), 2.73-2.78 (m, 1H), 2.40-2.52 (m, 5H), 2.08- 2.14
(m, 1H), 1.55-1.70 (m, 2H); LCMS: 95.5%, MS (ESI): m/z 590.3 [M +
H]+. 95 ##STR00303## white solid; .sup.1H-NMR (MeOD, 400 MHz):
.delta. 8.45 (d, J = 5.2 Hz, 1H), 8.34 (t, J = 8.8 Hz, 1H), 7.77
(s, 1H), 7.59 (s, 1H), 7.36 (s, 1H), 7.22 (dd, J = 11.2, 2.4 Hz,
1H), 7.12 (d, J = 8.8 Hz, 1H), 6.64 (d, J = 5.6 Hz, 1H), 4.84 (t, J
= 7.2 Hz, 2H), 4.64 (q, J = 8.8 Hz, 2H), 4.58 (t, J = 6.0 Hz, 2H),
4.50 (d, J = 7.2 Hz, 2H), 4.02 (s, 3H), 4.00 (s, 3H), 3.50-3.63 (m,
1H); LCMS: 100%, MS (ESI): m/z 577.1 [M + H]+. 96 ##STR00304##
yellow solid; .sup.1H-NMR (CDCl3, 400 MHz): .delta. 8.74 (s, 1H),
8.51 (d, J = 5.2 Hz, 1H), 7.89 (dd, J = 2.0, 12.0 Hz, 1H), 7.60 (s,
1H), 7.59 (s, 1H), 7.44 (s, 1H), 7.36-7.42 (m, 1H), 7.23-7.25 (m,
1H), 6.44 (d, J = 5.2 Hz, 1H), 4.86-4.90 (m, 1H), 4.55 (q, J = 8.4
Hz, 2H), 4.08 (s, 3H), 4.06 (s, 3H), 2.97-3.06 (m, 2H), 2.73- 2.77
(m, 1H), 2.40-2.52 (m, 5H), 2.12-2.16 (m, 1H), 1.55-1.73 (m, 2H);
LCMS: 98.3%, MS (ESI): m/z 590.3 [M + H]+. 97 ##STR00305## white
solid; .sup.1H-NMR (MeOD, 400 MHz): .delta. 8.43 (d, J = 5.2 Hz,
1H), 7.94 (dd, J = 12.8, 2.0 Hz, 1H), 7.74 (s, 1H), 7.64 (s, 1H),
7.55 (br d, J = 8.8 Hz, 1H), 7.33-7.39 (m, 2H), 6.52 (d, J = 5.6
Hz, 1H), 4.83 (t, J = 7.2 Hz, 2H), 4.54-4.65 (m, 4H), 4.49 (d, J =
7.6 Hz, 2H), 4.02 (s, 6H), 3.50-3.63 (m, 1H); LCMS: 100%, MS (ESI):
m/z 577.2 [M + H]+. 98 ##STR00306## yellow solid; .sup.1H-NMR
(CDCl3, 400 MHz): .delta. 8.64 (s, 1H), 8.45 (d, J = 5.6 Hz, 1H),
7.69 (d, J = 2.4 Hz, 1H), 7.62 (s, 1H), 7.57 (s, 1H), 7.54 (d, J =
2.0 Hz, 1H), 7.43 (s, 1H), 7.10 (d, J = 8.4 Hz, 1H), 6.30 (d, J =
5.2 Hz, 1H), 4.84-4.88 (m, 1H), 4.55 (q, J = 8.4 Hz, 2H), 4.07 (s,
3H), 4.05 (s, 3H), 2.96-3.01 (m, 2H), 2.74- 2.78 (m, 1H), 2.50-2.60
(m, 1H), 2.42-2.45 (m, 4H), 2.09-2.19 (m, 4H); LCMS: 97.0%, MS
(ESI): m/z 586.3 [M + H]+. 99 ##STR00307## white solid; .sup.1H-NMR
(MeOD, 400 MHz): .delta. 8.39 (d, J = 5.2 Hz, 1H), 7.74 (br s, 2H),
7.63-7.70 (m, 2H), 7.36 (s, 1H), 7.16 (d, J = 8.8 Hz, 1H), 6.39 (d,
J = 5.2 Hz, 1H), 4.84 (t, J = 7.2 Hz, 2H), 4.55-4.66 (m, 4H), 4.49
(d, J = 7.6 Hz, 2H), 4.02 (s, 6H), 3.50- 3.63 (m, 1H), 2.19 (s,
3H); LCMS: 100%, MS (ESI): m/z 573.2 [M + H]+. 100 ##STR00308##
yellow solid; .sup.1H-NMR (CDCl3, 400 MHz): .delta. 8.69 (s, 1H),
8.50 (d, J = 5.2 Hz, 1H), 7.77 (d, J = 8.8 Hz, 2H), 7.59 (d, J =
2.8 Hz, 2H), 7.43 (s, 1H), 7.20 (d, J = 8.4 Hz, 2H), 6.49 (d, J =
5.2 Hz, 1H), 4.86-4.91 (m, 1H), 4.56 (q, J = 8.0 Hz, 2H), 4.07 (s,
6H), 2.97-3.03 (m, 2H), 2.76-2.78 (m, 1H), 2.43-2.45 (m, 5H), 2.10-
2.15 (m, 1H), 1.55-1.70 (m, 2H); LCMS: 97.9%, MS (ESI): m/z 572.3
[M + H]+. 101 ##STR00309## white solid; .sup.1H-NMR (MeOD, 400
MHz): .delta. 8.42 (d, J = 5.6 Hz, 1H), 7.84 (s, 1H), 7.82 (s, 1H),
7.74 (s, 1H), 7.63 (s, 1H), 7.35 (s, 1H), 7.26 (s, 1H), 7.24 (s,
1H), 6.55 (d, J = 5.2 Hz, 1H), 4.84 (t, J = 7.2 Hz, 2H), 4.55-4.65
(m, 4H), 4.49 (d, J = 7.2 Hz, 2H), 4.01 (br s, 3H), 4.00 (br s,
3H), 3.50-3.63 (m, 1H); LCMS: 100%, MS (ESI): m/z 559.2 [M + H]+.
102 ##STR00310## white solid; .sup.1H-NMR (CDCl3, 400 MHz): .delta.
9.79 (s, 1H), 8.97 (d, J = 8.8 Hz, 1H), 8.60-8.63 (m, 1H), 8.23 (s,
1H), 7.58-7.61 (m 2H), 7.36-7.40 (m, 2H), 4.53 (q, J = 8.0 Hz, 2H),
4.21 (s, 3H), 4.12 (s, 3H), 3.98 (s, 3H); LCMS: 98.9%, MS (ESI):
m/z 505.2 [M + H]+. 103 ##STR00311## white solid; .sup.1H-NMR
(DMSO-d6, 400 MHz): .delta. 10.32 (s, 1H), 8.75-8.76 (m, 2H), 8.45
(dd, J = 2.0, 8.8 Hz, 1H), 7.90 (s, 1H), 7.60 (s, 1H), 7.55 (s,
1H), 7.43 (d, J = 9.2 Hz, 1H), 7.11 (d, J = 6.4 Hz, 1H), 4.68 (q, J
= 9.2 Hz, 2H), 4.00 (s, 3H), 3.96 (s, 3H), 3.93 (d, J = 7.2 Hz,
2H), 2.11-2.20 (m, 1H), 0.87 (s, 3H), 0.85 (s, 3H); LCMS: 96.8%, MS
(ESI): m/z 546.3 [M + H]+. 104 ##STR00312## white solid;
.sup.1H-NMR (MeOD, 400 MHz): .delta. 8.62 (s, 1H), 8.49- 8.53 (m,
1H), 8.36-8.38 (m, 1H), 7.69 (m, 1H), 7.49-7.51 (m, 1H), 7.35-7.38
(m, 1H), 7.29-7.30 (m, 1H), 6.88-6.89 (m, 2H), 4.59 (q, J = 8.8 Hz,
2H), 4.31-4.33 (m, 2H), 4.40 (d, J = 3.6 Hz, 3H), 3.95 (d, J = 4.0
Hz, 3H), 3.78-3.80 (m, 2H), 3.35 (s, 3H); LCMS: 99.7%, MS (ESI):
m/z 548.2 [M + H]+. 105 ##STR00313## yellow solid; .sup.1H-NMR
(MeOD, 400 MHz): .delta. 8.75 (d, J = 6.8 Hz, 1H), 8.53 (d, J = 2.4
Hz, 1H), 8.43 (d, J = 9.2 Hz, 1H), 8.10 (dd, J = 2.4, 9.2 Hz, 1H),
7.87 (s, 1H), 7.71 (s, 1H), 7.53 (s, 1H), 7.12 (d, J = 6.8 Hz, 1H),
4.24-4.27 (m, 2H), 4.15-4.16 (m, 5H), 4.10 (s, 3H), 3.81-3.83 (m,
2H), 3.45 (s, 3H), 1.95 (d, J = 7.2 Hz, 2H), 0.96 (t, J = 7.6 Hz,
3H); LCMS: 99.2%, MS (ESI): m/z 508.2 [M + H]+.
106 ##STR00314## yellow solid; .sup.1H-NMR (MeOD, 400 MHz): .delta.
8.46-8.48 (m, 2H), 8.33 (d, J = 2.8 Hz, 1H), 7.79 (dd, J = 2.8, 3.2
Hz, 1H), 7.73 (s, 1H), 7.66 (s, 1H), 7.39 (s, 1H), 6.61 (d, J = 5.2
Hz, 1H), 4.91- 4.95 (m, 2H), 4.67 (t, J = 6.0 Hz, 2H), 4.36 (d, J =
6.8 Hz, 2H), 4.16 (t, J = 6.8 Hz, 2H), 4.03 (d, J = 3.2 Hz, 6H),
3.52-3.68 (s, 1H), 1.92-1.98 (m, 2H), 0.97 (t, J = 7.2 Hz, 3H);
LCMS: 99.5%, MS (ESI): m/z 520.1 [M + H]+. 107 ##STR00315## yellow
solid; .sup.1H-NMR (MeOD, 400 MHz): .delta. 8.76 (d, J = 6.4 Hz,
1H), 8.53 (d, J = 1.8 Hz, 1H), 8.43 (d, J = 9.2 Hz, 1H), 8.10 (dd,
J = 2.4, 9.2 Hz, 1H), 7.87 (s, 1H), 7.76 (s, 1H), 7.54 (s, 1H),
7.11 (d, J = 6.8 Hz, 1H), 4.24 (dd, J = 4.4, 5.2 Hz, 2H), 4.16 (s,
3H), 4.12 (s, 3H), 3.79-3.83 (m, 3H), 3.47 (s, 3H), 1.22-1.26 (m,
2H), 1.10-1.13 (m, 2H); LCMS: 97.7%, MS (ESI): m/z 506.1 [M + H]+.
108 ##STR00316## yellow solid; .sup.1H-NMR (MeOD, 400 MHz): .delta.
8.74 (d, J = 6.8 Hz, 1H), 8.53 (d, J = 2.4 Hz, 1H), 8.42 (d, J =
9.2 Hz, 1H), 8.12 (dd, J = 9.2, 2.8 Hz, 1H), 7.85 (s, 1H), 7.73 (s,
1H), 7.53 (s, 1H), 7.11 (d, J = 6.8 Hz, 1H), 4.45 (quin, J = 6.0
Hz, 1H), 4.14 (s, 3H), 4.10 (s, 3H), 3.75-3.84 (m, 1H), 1.43 (s,
3H), 1.42 (s, 3H), 1.21-1.26 (m, 2H), 1.05-1.12 (m, 2H); LCMS:
98.1%, MS (ESI): m/z 490.2 [M + H]+. 109 ##STR00317## yellow solid;
.sup.1H-NMR (CDCl3, 400 MHz): .delta. 8.79 (d, J = 6.8 Hz, 1H),
8.63 (s, 1H), 8.30-8.37 (m, 2H), 7.87 (s, 1H), 7.84 (s, 1H), 7.57
(s, 1H), 7.20 (d, J = 6.8 Hz, 1H), 6.11-6.40 (m, 1H), 4.30- 4.37
(m, 2H), 4.16 (s, 3H), 4.12 (s, 3H), 3.80-3.86 (m, 1H), 3.32-3.34
(m, 1H), 1.26-1.29 (m, 2H), 1.11-1.15 (m, 2H); LCMS: 99.2%, MS
(ESI): m/z 512.3 [M + H]+. 110 ##STR00318## white solid;
.sup.1H-NMR (MeOD, 400 MHz): .delta. 8.74 (d, J = 6.4 Hz, 1H), 8.51
(s, 2H), 8.03 ( d, J = 9.2 Hz, 1H), 7.87 (s, 1H), 7.78 (s, 1H),
7.52 (s, 1H), 7.08 (d, J = 6.8 Hz, 1H), 4.86-4.88 (m, 1H),
4.74-4.76 (m, 1H), 4.38-4.40 (m, 1H), 4.31-4.32 (m, 1H), 4.15 (s, 3
H), 4.12 (s, 3H), 3.78-3.81 (m, 1H), 1.22-1.27 (m, 2H), 1.08-1.13
(m, 2H); LCMS: 99.1%, MS (ESI): m/z 494.2 [M + H]+. 111
##STR00319## yellow solid; .sup.1H-NMR (MeOD, 400 MHz): .delta.
8.74 (d, J = 6.4 Hz, 1H), 8.52 (br s, 1H), 8.41 (d, J = 7.2 Hz,
1H), 8.12 (d, J = 5.6 Hz, 1H), 7.84 (s, 1H), 7.71 (s, 1H), 7.53 (s,
1H), 7.11 (d, J = 6.4 Hz, 1H), 4.13 (s, 3H), 4.10 (s, 3H), 4.05 (d,
J = 6.4 Hz, 2H), 3.73- 3.80 (m, 1H), 2.80-2.93 (m, 1H), 2.11-2.24
(m, 2H), 2.00 (br s, 4H), 1.19-1.26 (m, 2H), 1.04-1.12 (m, 2H);
LCMS: 99.1%, MS (ESI): m/z 516.2 [(M + H)]+. 112 ##STR00320## white
solid; .sup.1H-NMR (MeOD, 400 MHz): .delta. 8.77 (d, J = 9.6 Hz,
1H), 8.54 (d, J = 5.6 Hz, 1H), 7.81 (s, 1H), 7.23(d, J = 9.6 Hz,
1H), 7.49 (s, 1H), 7.41 (s, 1H), 7.07 (d, J = 5.2 Hz, 1H), 4.62 (q,
J = 8.4 Hz, 1H), 4.03 (s, 3H), 3.96 (s, 3H), 3.75-3.79 (m, 1H),
1.20-1.24 (m, 2H), 1.05-1.10 (m, 2H); LCMS: 98.9%, MS (ESI): m/z
531.2 [(M + H)]+. 113 ##STR00321## yellow solid; .sup.1H-NMR
(CDCl3, 400 MHz): .delta. 9.75 (s, 1H), 8.86 (d, J = 9.6 Hz, 1H),
8.67 (d, J = 5.2 Hz, 1H), 7.51 (s, 1H), 7.42 (d, J = 9.2 Hz, 2H),
7.34 (s, 1H), 6.97 (d, J = 5.2 Hz, 1H), 4.86- 4.88 (m, 1H),
4.75-4.77 (m, 1H), 4.36-4.38 (m, 1H), 4.26-4.31 (m, 1H), 4.08 (s,
3H), 4.03 (s, 3H), 3.62-3.68 (m, 1H), 1.23- 1.27 (m, 2H), 1.05-1.14
(m, 2H); LCMS: 98.9%, MS (ESI): m/z 495.2 [(M + H)]+. 114
##STR00322## yellow solid; .sup.1H-NMR (MeOD, 400 MHz): .delta.
8.74 (d, J = 6.8 Hz, 1H), 8.45 (d, J = 2.4 Hz, 1H), 8.23 (dd, J =
8.8, 2.4 Hz, 1H), 7.82 (s, 1H), 7.76 (s, 1H), 7.60 (d, J = 8.8 Hz,
1H), 7.52 (s, 1H), 7.03 (d, J = 6.8 Hz, 1H), 4.63 (q, J = 8.4 Hz,
2H), 4.16-4.19 (m, 5H), 4.11 (s, 3H), 1.92-2.01 (m, 2H), 0.97 (t, J
= 7.6 Hz, 3H); LCMS: 98.7%, MS (ESI): m/z 599.2 [(M + H)]+. 115
##STR00323## white solid; .sup.1H-NMR (MeOD, 400 MHz): .delta. 8.69
(d, J = 6.8 Hz, 1H), 8.49 (t, J = 8.8 Hz, 1H), 7.83 (s, 1H), 7.74
(s, 1H), 7.48 (s, 1H), 7.41 (dd, J = 2.4,11.2 Hz, 1H), 7.27 (d, J =
9.2 Hz, 1H), 7.03 (d, J = 6.8 Hz, 1H), 4.65 (q, J = 8.4 Hz, 2H),
4.33 (t, J = 5.2 Hz, 2H), 4.13 (s, 3H), 4.09 (s, 3H), 3.92 (t, J =
5.2 Hz, 2H), 3.33-3.36 (m, 1H), 0.46-0.48 (m, 4H); LCMS: 95.6%, MS
(ESI): m/z 591.2 [(M + H)]+. 116 ##STR00324## off-white solid;
.sup.1H-NMR (MeOD, 400 MHz): .delta. 8.71 (d, J = 6.8 Hz, 1H), 8.06
(dd, J = 2.4,12.8 Hz, 1H), 7.85 (s, 1H), 7.70 (s, 1H), 7.65 (d, J =
8.8 Hz, 1H), 7.46-7.53 (m, 2H), 7.01 (d, J = 6.8 Hz, 1H), 4.61 (q,
J = 8.8 Hz, 2H), 4.33 (t, J = 4.8 Hz, 2H), 4.14 (s, 3H), 4.10 (s,
3H), 3.92 (t, J = 4.8 Hz, 2H), 3.34-3.37 (m, 1H), 0.46-0.48 (m,
4H); LCMS: 95.7%, MS (ESI); m/z 591.2 [(M + H)]+. 117 ##STR00325##
light yellow solid; .sup.1H-NMR (MeOD, 400 MHz): .delta. 8.65 (d, J
= 6.8 Hz, 1H), 7.88 (s, 1H), 7.83 (d, J = 2.0 Hz, 1H), 7.76 (dd, J
= 2.4.8.8 Hz, 1H), 7.70 (s, 1H), 7.48 (s, 1H), 7.27 (d, J = 8.4 Hz,
1H), 6.84 (d, J = 6.8 Hz, 1H), 4.61 (q, J = 8.4 Hz, 2H), 4.32 (t, J
= 5.2 Hz, 2H), 4.13 (s, 3H), 4.11 (s, 3H), 3.92 (t, J = 5.2 Hz,
2H), 3.33-3.37 (m, 1H), 2.23 (s, 3H), 0.44-0.50 (m, 4H); LCMS:
96.7%, MS (ESI): m/z 587.2 [(M + H)]+. 118 ##STR00326## yellow
solid; .sup.1H-NMR (MeOD, 400 MHz): .delta. 8.67 (d, J = 6.8 Hz,
1H), 7.94 (d, J = 8.8 Hz, 2H), 7.84 (s, 1H), 7.70 (s, 1H), 7.49 (s,
1H), 7.37 (d, J = 8.8 Hz, 2H), 6.95 (d, J = 6.8 Hz, 1H), 4.62 (q, J
= 8.8 Hz, 2H), 4.33 (t, J = 5.2 Hz, 2H), 4.13 (s, 3H), 4.10 (s,
3H), 3.92 (t, J = 5.2 Hz, 2H), 3.31-3.35 (m, 1H), 0.46-0.49 (m,
4H); LCMS: 97.0%, MS (ESI): m/z 573.2 [(M + H)]+. 119 ##STR00327##
HCl salt, yellow solid; 1H-NMR (MeOD, 400 MHz): .delta. 8.68 (d, J
= 6.8 Hz, 1H), 7.97 (d, J = 8.8 Hz, 2H), 7.87 (s, 1H), 7.83 (s,
1H), 7.51 (s, 1H), 7.40 (d, J = 8.8 Hz, 2H), 6.95 (d, J = 6.8 Hz,
1H), 4.66 (q, J = 8.4 Hz, 2H), 4.60 (t, J = 4.8 Hz, 2H), 4.12 (s,
3H), 4.09 (s, 3H), 3.63 (t, J = 4.8 Hz, 2H), 2.83 (s, 3H); LCMS:
94.0%, MS (ESI): m/z 546.2 [(M + H)]+. 120 ##STR00328## off-white
solid; .sup.1H-NMR (MeOD, 400 MHz): .delta. 8.64 (d, J = 9.6 Hz,
1H), 8.45 (d, J = 5.6 Hz, 1H), 7.59 (t, J = 4.4 Hz, 2H), 7.36 (s,
1H), 7.28 (s, 1H), 6.94 (d, J = 5.6 Hz, 1H), 4.51 (q, J = 8.4 Hz,
2H), 4.20 (t, J = 5.2 Hz, 2H), 3.90 (s, 3H), 3.84 (s, 3H), 3.79 (t,
J = 5.2 Hz, 2H), 3.21-3.24 (m, 1H), 0.33-0.35 (m, 4H); LCMS: 98.6%,
MS (ESI): m/z 575.2 [(M + H)]+. 121 ##STR00329## yellow solid;
.sup.1H-NMR (MeOD, 400 MHz): .delta. 8.90 (d, J = 9.2 Hz, 1H), 8.79
(d, J = 6.8 Hz, 1H), 7.89 (d, J = 9.6 Hz, 1H), 7.82 (d, J = 8.4 Hz,
1H), 7.55 (d, J = 6.8 Hz, 1H), 7.49 (s, 1H), 4.21-4.77 (m, 1H),
4.65 (dd, J = 8.4 Hz, 2H), 4.14 (s, 3H), 4.09 (s, 3H), 2.09-2.21
(m, 2H), 2.08-2.09 (m, 2H), 1.93-1.94 (m,2H), 1.76- 1.77 (m, 2H);
LCMS: 93.2%, MS (ESI): m/z 559.2 [M + H]+. 122 ##STR00330## HCl
salt, yellow solid; .sup.1H-NMR (MeOD, 400 MHz): .delta. 8.91 (d, J
= 9.6 Hz, 1H), 8.82 (d, J = 6.8 Hz, 1H, 7.89-7.97 (m, 2H), 7.84 (s,
1H), 7.56 (d, J = 6.4 Hz, 1H), 7.52 (s, 1H), 5.31 (t, J = 6.0 Hz,
1H ), 4.69 (q, J = 8.4 Hz, 2H), 4.15 (s, 3H), 4.10 (s, 3H), 3.93
(br d, J = 13.2 Hz, 1H), 3.63-3.76 (m, 2H), 3.51-3.60 (m, 1H),
2.54-2.73 (m, 1H), 2.34-2.44 (m, 1H); LCMS: 97.5%, MS (ESI): m/z
560.2 [M + H]+. 123 ##STR00331## white solid; .sup.1H-NMR (MeOD,
400 MHz): .delta. 8.78 (d, J = 9.6 Hz, 1H), 8.60 (d, J = 5.2 Hz,
1H), 7.80 (s, 1H), 7.74 (d, J = 9.6 Hz, 1H), 7.50 (s, 1H), 7.43 (s,
1H), 7.08 (d, J = 5.2 Hz, 1H), 4.84- 4.86 (m, 2H), 4.63 (q, J = 8.4
Hz, 2H), 4.59 (t, J = 6.4 Hz, 2H), 4.52 (d, J = 7.6 Hz, 2H), 4.05
(s, 3H), 3.98 (s, 3H), 3.53-3.63 (m, 1H); LCMS: 96.0%, MS (ESI):
m/z 561.0 [M + H]+. 124 ##STR00332## yellow solid; .sup.1H-NMR
(MeOD, 400 MHz): .delta. 8.85 (d, J = 9.2 Hz, 1H), 8.82 (d, J = 6.8
Hz, 1H), 7.97 (d, J = 9.6 Hz, 1H), 7.83 (s, 1H), 7.78 (s, 1H), 7.60
(d, J = 6.8 Hz, 1H), 7.52 (s, 1H), 4.65 (q, J = 8.4 Hz, 2H), 4.14
(s, 3H), 4.09 (s, 3H), 4.01 (d, J = 7.2 Hz, 2H), 2.21-2.31 (m, 1H),
0.96 (s, 3H), 0.94 (s, 3H); LCMS: 98.6%, MS (ESI): m/z 547.2 [M +
H]+. 125 ##STR00333## HCl salt, white solid; .sup.1H-NMR (MeOD, 400
MHz): .delta. 8.89 (d, J = 9.6 Hz, 1H), 8.85 (d, J = 6.4 Hz, 1H),
8.07 (d, J = 9.2 Hz, 1H), 7.89 (s, 1H), 7.83 (s, 1H), 7.65 (d, J =
6.8 Hz, 1H), 7.55 (s, 1H), 4.66 (q, J = 8.4 Hz, 2H), 4.55 (t, J =
5.2 Hz, 2H), 4.15 (s, 3H), 4.09 (s, 3H), 3.56 (d, J = 4.8 Hz, 2H);
LCMS: 96.3%, MS (ESI): m/z 534.2 [M + H]+. 126 ##STR00334## yellow
solid; .sup.1H-NMR (MeOD, 400 MHz): .delta. 8.80-8.83 (m, 2H), 8.00
(d, J = 9.26 Hz, 1H), 7.81 (s, 1H), 7.78 (s, 1H), 7.61 (d, J = 6.8
Hz, 1H), 7.55 (s, 1H), 4.64 (q, J = 8.4 Hz, 2H), 4.37 (t, J = 4.8
Hz, 2H), 4.14 (s, 3H), 4.09 (s, 3H), 3.81 (t, J = 4.8 Hz, 2H), 3.36
(s, 3H); LCMS: 93.4%, MS (ESI): m/z 549.2 [M + H]+. 127
##STR00335## HCl salt, off-white solid; .sup.1H-NMR (MeOD, 400
MHz): .delta. 8.88 (d, J = 9.6 Hz, 1H), 8.82 (d, J = 6.4 Hz, 1H),
7.96 (d, J = 9.6 Hz, 1H), 7.91 (s, 1H), 7.82 (s, 1H), 7.57 (d, J =
6.8 Hz, 1H), 7.55 (s, 1H), 4.68 (q, J = 8.4 Hz, 2H), 4.61 (t, J =
4.8 Hz, 2H), 4.15 (s, 3H), 4.09 (s, 3H), 3.64 (t, J = 4.8 Hz, 2H),
2.83 (s, 3H); LCMS: 98.3%, MS (ESI): m/z 548.2 [(M + H)]+. 128
##STR00336## off-white solid; .sup.1H-NMR (MeOD, 400 MHz): .delta.
8.87 (d, J = 9.6 Hz, 1H), 8.81 (d, J = 6.4 Hz, 1H), 7.91 (d, J =
9.6 Hz, 1H), 7.84 (s, 1H), 7.83 (s, 1H), 7.56 (d, J = 6.4 Hz, 1H),
7.51 (s, 1H), 5.08- 5.10 (m, 1H), 4.64 (q, J = 8.4 Hz, 2H),
4.12-4.22 (m, 5H), 4.09 (s, 3H), 4.01-4.05 (m, 1H), 3.91-3.93 (m,
1H), 2.53-2.56 (m, 1H), 2.37-2.42 (m, 1H); LCMS: 98.6%, MS (ESI):
m/z 561.2 [M + H]+. 129 ##STR00337## white solid; .sup.1H-NMR
(MeOD, 400 MHz): .delta. 8.79 (d, J = 2.4 Hz, 1H), 8.73 (d, J = 6.8
Hz, 1H), 8.48 (dd, J = 2.8, 8.8 Hz, 1H), 7.81 (s, 1H), 7.73 (s,
1H), 7.47-7.49 (m, 2H), 7.33 (d, J = 6.8 Hz, 1H), 4.61 (q, J = 8.8
Hz, 2H), 4.27 (t, J = 5.2 Hz, 2H), 4.13 (s, 3H), 4.09 (s, 3H), 3.96
(t, J = 5.2 Hz, 2H); LCMS: 100%, MS (ESI): m/z 534.2 [M + H]+. 130
##STR00338## off-white solid; .sup.1H-NMR (MeOD, 400 MHz): .delta.
8.63 (d, J = 2.4 Hz, 1H), 8.52 (d, J = 5.2 Hz, 1H), 8.38 (dd, J =
2.8, 8.8 Hz, 1H), 7.67 (s, 1H), 7.52 (s, 1H), 7.38 (s, 1H), 7.30
(d, J = 8.8 Hz, 1H), 6.89 (d, J = 5.6 Hz, 1H), 4.59 (q, J = 8.4 Hz,
2H), 4.31 (t, J = 5.2 Hz, 2H), 4.02 (s, 3H), 3.97 (s, 3H), 3.90 (t,
J = 5.2 Hz, 2H), 3.33-3.34 (m, 1H), 0.45-0.47 (m, 4H); LCMS: 98.6%,
MS (ESI): m/z 574.2 [(M + H)]+. 131 ##STR00339## white solid;
.sup.1H-NMR (MeOD, 400 MHz): .delta. 8.79 (d, J = 2.8 Hz, 1H), 8.73
(d, J = 6.8 Hz, 1H), 8.48 (dd, J = 2.8, 8.8 Hz, 1H), 7.82 (s, 1H),
7.77 (s, 1H), 7.46-7.49 (m, 2H), 7.33 (d, J = 7.2 Hz, 1H),
4.72-4.76 (m, 1H), 4.58-4.64 (m, 2H), 4.13 (s, 3H), 4.09 (s, 3H),
2.19-2.22 (m, 2H), 2.08-2.11 (m, 2H), 1.92-1.93 (m, 2H), 1.75-1.78
(m, 2H); LCMS: 100%, MS (ESI): m/z 558.2 [M + H]+. 132 ##STR00340##
HCl salt, yellow solid; .sup.1H-NMR (MeOD, 400 MHz): .delta. 8.89
(d, J = 2.4 Hz, 1H), 8.72 (d, J = 6.8 Hz, 1H), 8.56 (dd, J = 2.4,
8.8 Hz, 1H), 7.89 (s, 1H), 7.74 (s, 1H), 7.45-7.48 (m, 2H), 7.33
(d, J = 6.8 Hz, 1H), 5.26-5.33 (m, 1H), 4.62-4.64 (m, 2H), 4.11 (s,
3H), 4.07 (s, 3H), 3.99 (d, J = 12.8 Hz, 1H), 3.71-3.76 (m, 2H),
3.53- 3.59 (m, 1H), 2.59-2.62 (m, 1H), 2.39-2.41 (m, 1H); LCMS:
96.1%, MS (ESI): m/z 559.2 [M + H]+. 133 ##STR00341## yellow solid;
.sup.1H-NMR (DMSO-d6, 400 MHz): .delta. 11.02 (s, 1H), 9.13 (d, J =
2.4 Hz, 1H), 8.87 (d, J = 6.4 Hz, 1H), 8.76 (dd, J = 2.8, 9.2 Hz,
1H), 8.03 (s, 1H), 7.74 (s, 1H), 7.69 (s, 1H), 7.53 (d, J = 8.8 Hz,
1H), 7.26 (d, J = 6.4 Hz, 1H), 5.29-5.32 (m, 1H), 4.70 (q, J = 9.2
Hz, 2H), 4.05 (s, 3H), 4.04 (s, 3H), 3.78-3.85 (m, 2H), 3.16-3.23
(m, 2H), 2.90-2.91 (m, 3H), 2.74-2.77 (m, 1H), 2.17-2.22 (m, 1H);
LCMS: 95.4%, MS (ESI): m/z 573.3 [M + H]+. 134 ##STR00342## white
solid; .sup.1H-NMR (MeOD, 400 MHz): .delta. 8.61 (d, J = 2.8 Hz,
1H), 8.51 (d, J = 5.6 Hz, 1H), 8.36 (dd, J = 2.8, 8.8 Hz, 1H), 7.73
(s, 1H), 7.50 (s, 1H), 7.38 (s, 1H), 7.30 (d, J = 8.8 Hz, 1H), 6.88
(d, J = 5.2 Hz, 1H), 4.83 (t, J = 6.8 Hz, 2H), 4.55-4.62 (m, 4H),
4.48 (d, J = 7.6 Hz, 2H), 4.01 (s, 3H), 3.96 (s, 3H), 3.52-3.61 (m,
1H).; LCMS: 98.4%, MS (ESI): m/z 560.2 [M + H]+. 135 ##STR00343##
HCl salt, yellow solid; .sup.1H-NMR (MeOD, 400 MHz): .delta. 8.88
(d, J = 5.2 Hz, 1H), 8.80 (d, J = 2.4 Hz, 1H), 8.73 (d, J = 6.8 Hz,
1H), 8.50 (dd, J = 2.4, 8.8 Hz, 1H), 8.13 (t, J = 6.8 Hz, 1H), 7.83
(s, 1H), 7.81 (s, 1H), 7.50 (d, J = 8.8 Hz, 1H), 7.48 (s, 1H), 7.35
(d, J = 6.8 Hz, 1H), 4.64 (q, J = 8.8 Hz, 2H), 4.51 (t, J = 6.4 Hz,
2H), 4.13 (s, 3H), 4.08 (s, 3H), 3.54 (t, J = 5.6 Hz, 2H); LCMS:
93.6%, MS (ESI): m/z 533.2 [M + H]+. 136 ##STR00344## yellow solid;
.sup.1H-NMR (MeOD, 400 MHz): .delta. 8.79 (d, J = 2.4 Hz, 1H), 8.72
(d, J = 6.4 Hz, 1H), 8.48 (dd, J = 2.8, 8.8 Hz, 1H), 7.81 (s, 1H),
7.71 (s, 1H), 7.47-7.49 (m, 2H), 7.33 (d, J = 6.8 Hz, 1H), 4.60 (q,
J = 8.8 Hz, 2H), 4.18-4.21 (m, 2H), 4.13 (s, 3H), 4.03-4.08 (m,
4H), 1.20 (d, J = 6.0 Hz, 3H); LCMS: 98.9%, MS (ESI): m/z 548.2 [M
+ H]+. 137 ##STR00345## HCl salt, off-white solid; .sup.1H-NMR
(MeOD, 400 MHz): .delta. 8.83 (d, J = 2.4 Hz, 1H), 8.73 (d, J = 6.8
Hz, 1H), 8.52 (dd, J = 8.8, 2.4 Hz, 1H), 7.85 (s, 1H), 7.78 (s,
1H), 7.48-7.51 (m, 2H), 7.35 (d, J = 6.4 Hz, 1H), 4.64 (q, J = 8.8
Hz, 2H), 4.59 (t, J = 5.6 Hz, 2H), 4.13 (S, 3H), 4.08 (s, 3H), 3.63
(t, J = 5.2 Hz, 2H), 2.83 (s, 3H); LCMS: 96.6%, MS (ESI): m/z 547.2
[(M + H)]+. 138 ##STR00346## yellow solid; .sup.1H-NMR (MeOD, 400
MHz): .delta. 8.79 (d, J = 2.8 Hz, 1H), 8.73 (d, J = 6.8 Hz, 1H),
8.48 (dd, J = 8.4, 2.4 Hz, 1H), 7.78 (s, 1H), 7.81 (s, 1H),7.47 (t,
J = 4.8 Hz, 2H), 7.34 (d, J = 6.4 Hz, 1H), 5.06-5.09 (m, 1H),
4.60-4.62 (m, 2H), 4.17-4.19 (m, 2H), 4.13 (s, 3H), 4.09 (s, 3H),
4.02-4.04 (m, 1H), 3.91-3.93 (m, 1H), 2.50-2.57 (m, 1H), 2.37-2.40
(m, 1H); LCMS: 96.1%, MS (ESI): m/z 560.0 [M + H]+. 139
##STR00347## white solid; .sup.1H-NMR (MeOD, 400 MHz): .delta. 8.79
(d, J = 2.4 Hz, 1H), 8.73 (d, J = 6.8 Hz, 1H), 8.49 (dd, J = 8.4,
3.2 Hz, 1H), 7.81 (s, 1H), 7.74 (s, 1H), 7.47-7.49 (m, 2H), 7.33
(d, J = 7.2 Hz, 1H), 4.60 (q, J = 8.4 Hz, 2H), 4.14-4.16 (m, 2H),
4.13 (s, 3H), 4.09 (s, 3H), 1.93-1.95 (m, 2H), 0.95 (t, J = 7.2 Hz,
3H); LCMS: 99.5%, MS (ESI): m/z 532.2 [M + H]+. 140 ##STR00348##
HCl salt, yellow solid; .sup.1H-NMR (MeOD, 400 MHz): .delta. 8.71
(s, 1H), 8.61 (s, 1H), 8.29 (s, 2H), 7.75 (d, J = 9.2 Hz, 2H), 7.49
(s, 1H), 7.14 (s, 1H), 4.27 (s, 2H), 4.12 (t, J = 6.8 Hz, 2H), 4.05
(s, 3H), 4.02 (s, 3H), 3.41 ( s, 2H), 2.78 (s, 3H), 1.85-1.90 (m,
2H), 0.87 (t, J = 7.2 Hz, 3H); LCMS: 99.8%, MS (ESI): m/z 507.3 [M
+ H]+.
141 ##STR00349## HCl salt, yellow solid; .sup.1H-NMR (MeOD, 400
MHz): .delta. 8.71 (d, J = 6.4 Hz, 1H), 8.49 (d, J = 6.4 Hz, 1H),
8.48 (s, 1H), 7.98 (dd, J = 9.2, 2.8 Hz, 1H), 7.85 (s, 1H), 7.81
(s, 1H), 7.49 (s, 1H), 7.04 (d, J = 6.4 Hz, 1H), 4.29 (t, J = 4.8
Hz, 2H), 4.13 (s, 3H), 4.09 (s, 3H), 3.77-3.83 (m, 1H), 3.46 (t, J
= 4.8 Hz, 2H), 2.84 (s, 3H), 1.22-1.26 (m, 2H), 1.08-1.13 (m, 2H);
LCMS: 94.9%, MS (ESI): m/z 505.3 [M + H]+. 142 ##STR00350## yellow
solid; .sup.1H-NMR (MeOD, 400 MHz): .delta. 8.72 (d, J = 6.4 Hz,
1H), 8.46-8.48 (m, 2H), 8.00 (dd, J = 3.2, 9.2 Hz, 1H), 7.86 (s,
1H), 7.70 (S, 1H), 7.48 (s, 1H), 7.06 (d, J = 6.8 Hz, 1H), 4.14 (s,
3H), 4.10 (s, 3H), 3.82-3.84 (m, 1H), 3.52 (t, J = 7.2 Hz, 2H),
2.60-2.66 (m, 2H), 1.25-1.27 (m, 2H), 1.10-1.12 (m, 2H); LCMS:
98.4%, MS (ESI): m/z 543.2 [M + H]+. 143 ##STR00351## white solid;
.sup.1H-NMR (DMSO-d6, 400 MHz): .delta. 9.54 (s, 1H), 8.50 (d, J =
4.8 Hz, 1H), 8.38 (d, J = 2.4 Hz, 1H), 8.32 (d, J = 9.2 Hz, 1H),
7.95 (s, 1H), 7.82-7.89 (m, 1H), 7.54 (s, 1H), 7.42 (s, 1H), 6.55
(d, J = 5.2 Hz, 1H), 4.71-4.74 (m, 2H), 4.46 (t, J = 6.0 Hz, 2H),
4.28 (d, J = 6.4 Hz, 2H), 3.95 (d, J = 3.6 Hz, 6H), 3.78- 3.87 (m,
1H), 3.39-3.55 (m, 1H), 1.13-1.14 (m, 2H), 1.02-1.04 (m, 2H); LCMS:
99.3%, MS (ESI): m/z 518.1 [M + H]+. 144 ##STR00352## HCl salt,
yellow solid; .sup.1H-NMR (MeOD, 400 MHz): .delta. 8.76 (d, J = 6.8
Hz, 1H), 8.58 (s, 1H), 8.42 (d, J = 9.2 Hz, 1H), 8.18 (dd, J = 8.8,
2.0 Hz, 1H), 7.84 (d, J = 7.6 Hz, 2H), 7.55 (s, 1H), 7.14 (d, J =
6.8 Hz, 1H), 4.29-4.34 (m, 2H), 4.20-4.24 (m, 4H), 4.14 (s, 3H),
4.10 (s, 3H), 3.81-3.84 (m, 1H), 3.37-3.38 (m, 1H), 1.24-1.28 (m,
2H), 1.11-1.14 (m, 2H); LCMS: 97.0%, MS (ESI): m/z 517.2 [M + H]+.
145 ##STR00353## yellow solid; .sup.1H-NMR (CDCl3, 400 MHz):
.delta. 9.69-9.70 (m, 1H), 8.64-8.66 (m, 1H), 8.54-8.57 (m, 1H),
8.34 (s, 1H), 8.13 (s, 1H), 7.72-7.74 (m, 1H), 7.65 (s, 1H), 7.08
(s, 1H); 6.78 (br s, 1H), 4.19 (s, 3H), 4.15 (s, 3H), 3.60-3.68 (m,
3H), 1.24-1.28 (m, 2H); 1.06-1.08 (m, 2H); LCMS: 98.4%, MS (ESI):
m/z 529.2 [M + H]+. 146 ##STR00354## yellow solid; .sup.1H-NMR
(CDCl3, 400 MHz): .delta. 9.58 (d, J = 7.6 Hz, 1H), 8.79 (d, J =
6.8 Hz, 1H), 8.63 (d, J = 5.6 Hz, 1H), 7.61 (s, 1H), 7.49 (s, 1H),
7.43 (d, J = 9.6 Hz, 1H), 7.01-7.02 (m, 2H), 5.21 - 5.37 (m, 1H),
4.10 (s, 3H), 4.04 (s, 3H), 3.59-3.63 (m, 1H), 3.33 (t, J = 6.8 Hz,
2H), 2.43-2.50 (m, 2H), 1.20-1.24 (m, 2H), 1.04-1.09 (m, 2H); LCMS:
99.3%, MS (ESI): m/z 544.2 [M + H]+. 147 ##STR00355## white solid;
.sup.1H-NMR (MeOD, 400 MHz): .delta. 8.79 (d, J = 9.6 Hz, 1H),
8.53-8.58 (m, 1H), 7.71-7.73 (m, 2H), 7.48 (s, 1H), 7.40 (s, 1H),
7.05 (d, J = 5.2 Hz, 1H), 4.46 (q, J = 6.0 Hz, 1H), 4.02 (s, 3H),
3.96 (s, 3H), 3.73-3.77 (m, 1H), 1.43 (s, 3H), 1.42 (s, 3H),
1.19-1.22 (m, 2H), 1.06-1.09 (m, 2H).; LCMS: 99.1%, MS (ESI): m/z
491.2 [M + H]+. 148 ##STR00356## yellow solid; .sup.1H-NMR (CDCl3,
400 MHz): .delta. 9.50 (s, 1H), 8.70 (d, J = 9.6 Hz, 1H), 8.57 (d,
J = 5.6 Hz, 1H), 7.50 (s, 1H), 7.40 (s, 1H), 7.35 (d, J = 9.2 Hz,
1H), 7.00 (s, 1H), 6.93 (d, J = 5.2 Hz, 1H), 5.54 (br t, J = 6.8
Hz, 1H), 4.01 (s, 3H), 3.96 (s, 3H), 3.51-3.58 (m, 3H), 1.11-1.16
(m, 2H), 0.96-1.01 (m, 2H); LCMS: 97.6%, MS (ESI): m/z 530.2 [M +
H]+. 149 ##STR00357## yellow solid; .sup.1H-NMR (MeOD, 400 MHz):
.delta. 8.91 (d, J = 9.6 Hz, 1H), 8.79 (d, J = 6.8 Hz, 1H), 7.87
(d, J = 9.2 Hz, 1H), 7.83 (s, 1H), 7.73 (s, 1H), 7.54(d, J = 6.8
Hz, 1H), 7.50 (s, 1H), 4.14 (s, 3H), 4.07-4.09 (m, 5H), 3.74-3.79
(m, 1H), 2.88-2.91 (m, 1H), 2.16-2.21 (m, 2H), 1.99-2.02 (m, 4H),
1.20-1.24 (m, 2H), 1.07- 1.10 (m, 2H); LCMS: 99.4%, MS (ESI): m/z
517.3 [(M + H)]+. 150 ##STR00358## white solid; .sup.1H-NMR (CDCl3,
400 MHz): .delta. 9.50 (s, 1H), 8.55 (d, J = 9.2 Hz, 1H), 8.52 (d,
J = 5.2 Hz, 1H), 8.26 (d, J = 2.8 Hz, 1H), 7.59 (dd, J = 3.2, 9.2
Hz, 1H), 7.57 (s, 1H), 7.44 (s, 1H), 7.34 (s, 1H), 6.47 (d, J = 5.2
Hz, 1H), 4.30 - 4.33 (m, 1H), 4.20- 4.24 (m, 2 H), 4.18 (d, J = 2.8
Hz, 1H), 4.07 (d, J = 2.0 Hz, 6H), 3.97 - 4.05 (m, 1H), 3.83 (dd, J
= 3.2, 4.8 Hz, 2H), 3.48 (s, 3H), 2.42 (br s, 1H), 1.28 (d, J = 6.4
Hz, 3H); LCMS: 97.6%, MS (ESI): m/z 524.2 [(M + H)]+. 151
##STR00359## white solid; .sup.1H-NMR (CDCl3, 400 MHz): .delta.
9.59 (s, 1H), 8.56 (d, J = 8.8 Hz, 1H), 8.50 (d, J = 5.2 Hz, 1H),
8.25 (d, J = 2.8 Hz, 1H), 7.57-7.60 (m, 1H), 7.56 (s, 1H), 7.44 (s,
1H), 7.26 (s, 1H), 6.46 (d, J = 5.2 Hz, 1H), 4.31-4.36 (m, 2H),
4.20-4.23 (m, 1H), 4.07 (d, 6H), 3.04-4.06 (m, 1H), 2.66 (br, s,
1H), 1.47 (d, 6H), 1.28 (d, J = 6.0 Hz, 3H); LCMS: 99.6%, MS (ESI):
m/z 508.2 [(M + H)]+. 152 ##STR00360## white solid; .sup.1H-NMR
(DMSO-d6, 400 MHz) .delta. 9.64 (s, 1H), 8.85 (d, J = 6.8 Hz, 1H),
8.52 (d, J = 2.8 Hz, 1H), 8.42 (d, J = 9.2 Hz, 1H), 8.02 (dd, J =
9.2, 2.8 Hz, 1H), 7.80 (d, J = 6.4 Hz, 2H), 7.74 (s, 1H), 7.06 (d,
J = 6.8 Hz, 1H), 4.83 (t, J =3.6 Hz 1H), 4.71 (t, J = 3.6 Hz, 1H),
4.28-4.33 (m, 2H), 4.26-4.28 (m, 2H), 4.05 (s, 6H), 4.02-4.03 (m,
1H), 1.08 (d, J = 5.2 Hz, 3H); LCMS: 99.1%, MS (ESI): m/z 512.2 [M
+ H]+. 153 ##STR00361## off-white solid; .sup.1H-NMR (MeOD, 400
MHz): .delta. 8.74 (d, J = 6.8 Hz, 1H), 8.52 (s, 1H), 8.41 (d, J =
9.2 Hz, 1H), 8.10 (dd, J = 2.4, 8.8 Hz, 1H), 7.84 (s, 1H), 7.75 (s,
1H), 7.54 (s, 1H), 7.10 (d, J = 6.4 Hz, 1H), 4.64 (q, J = 8.8 Hz,
2H), 4.35 (t, J = 4.8 Hz, 2H), 4.13 (s, 3H), 4.10 (s, 3H), 3.93 (t,
J = 5.2 Hz, 2H), 3.30-3.36 (m, 1H), 0.46-0.47 (m, 4H); LCMS: 97.9%,
MS (ESI): m/z 574.2 [(M + H)]+. 154 ##STR00362## HCl salt, white
solid; .sup.1H-NMR (MeOD, 400 MHz): .delta. 8.46 (d, J = 5.2 Hz,
1H), 8.34 (t, J = 8.8 Hz, 1H), 7.76 (s, 1H), 7.61 (s, 1H), 7.37 (s,
1H), 7.22 (dd, J = 11.2, 2.4 Hz, 1H), 7.13 (dd, J = 8.8, 1.6 Hz,
1H), 6.65 (d, J = 5.6 Hz, 1H), 4.65 (q, J = 8.4 Hz, 2H), 4.30 (t, J
= 6.0 Hz, 2H), 4.02 (s, 1H), 4.00 (s, 1H), 3.07 (t, J = 6.0 Hz,
2H), 2.42 (s, 3H); LCMS: 100%, MS (ESI): m/z 564.2 [(M + H)]+. 155
##STR00363## HCl salt, yellow solid; .sup.1H-NMR (MeOD, 400 MHz):
.delta. 8.72 (d, J = 6.8 Hz, 1H), 8.09 (dd, J = 12.8, 2.4 Hz, 1H),
7.85 (s, 1H), 7.84 (s, 1H),7.68 (d, J = 8.8 Hz, 1H), 7.54 (t, J =
8.4 Hz, 1H), 7.50 (s, 1H), 7.02 (d, J = 6.0 Hz, 1H), 4.65 (q, J =
8.8 Hz, 2H), 4.57 (t, J = 5.2 Hz, 2H), 4.14 (s, 3H), 4.11 (s, 3H),
3.62 (t, J = 5.2 Hz, 2H), 2.82 (s, 3H); LCMS: 98.9%, MS (ESI): m/z
564.2 [(M + H)]+. 156 ##STR00364## HCl salt, off-white solid;
.sup.1H-NMR (MeOD, 400 MHz): .delta. 8.66 (d, J = 6.8 Hz, 1H), 7.88
(s, 1H), 7.85 (s, 1H), 7.84 (s, 1H), 7.80 (d, J = 8.4Hz, 1H), 7.51
(s, 1H), 7.30 (d, J = 8.8 Hz, 1H), 6.84 (d, J = 6.8 Hz, 1H), 4.66
(q, J = 8.8 Hz, 2H), 4.57 (t, J = 4.8 Hz, 2H), 4.14 (s, 3H), 4.11
(s, 3H), 3.62 (t, J = 4.8 Hz, 2H), 2.82 (s, 3H), 2.24 (s, 3H);
LCMS: 99.1%, MS (ESI): m/z 560.2 [(M + H)]+. 157 ##STR00365##
off-white solid; .sup.1H-NMR (MeOD, 400 MHz): .delta. 8.74 (d, J =
9.6 Hz, 1H), 8.56 (d, J = 5.6 Hz, 1H), 7.79 (s, 1H), 7.69 (d, J =
9.2 Hz, 1H), 7.47 (s, 1H), 7.39 (s, 1H), 7.04 (d, J = 5.6 Hz, 1H),
4.94-4.99 (m, 1H), 4.61 (q, J = 8.8 Hz, 2H), 4.01 (s, 3H), 3.95 (s,
3H), 3.06-3.09 (m, 2H), 2.81-2.85 (m, 1H), 2.45-2.52 (m, 5H),
2.27-2.28 (m, 1H); LCMS: 97.8%, MS (ESI): m/z 574.2 [M + H]+. 158
##STR00366## yellow solid; .sup.1H-NMR (MeOD, 400 MHz): .delta.
8.71 (d, J = 6.4 Hz, 1H), 8.50 (d, J = 9.2 Hz, 1H), 8.45 (d, J =
2.8 Hz, 1H), 7.94-7.97 (dd, J = 2.8, 9.2 Hz, 1H), 7.87 (s, 1H),
7.55 (s, 1H), 7.46 (s, 1H), 7.05 (d, J = 6.8 Hz, 1H), 4.17 (t, J =
7.2 Hz, 2H), 4.14 (s, 3H), 4.10 (s, 3H), 3.47 (t, J = 6.8 Hz, 2H),
2.58-2.61 (m, 2H), 1.93- 1.98 (m, 2H), 0.97 (t, J = 7.2 Hz, 3H);
LCMS: 98.0%, MS (ESI): m/z 545.2 [M + H]+. 159 ##STR00367## HCl
salt, yellow solid; .sup.1H-NMR (D.sub.2O, 400 MHz): .delta. 8.47
(d, J = 6.8 Hz, 1H), 8.22 (d, J = 2.8 Hz, 1H), 8.05 (d, J = 8.8 Hz,
1H), 7.72-7.81 (m, 1H), 7.54 (d, J = 7.6 Hz, 2H), 7.30 (s, 1H),
6.90 (d, J = 6.8 Hz, 1H), 4.20 (t, J = 9.6 Hz, 2H), 4.12-4.14 (m,
4H), 3.98-4.00 (m, 2H), 3.96 (s, 3H), 3.80 (s, 3H), 3.28-3.47 (m,
1H), 1.72-1.74 (m, 2H), 0.74 (t, J = 7.6 Hz, 3H); LCMS: 95.6%, MS
(ESI): m/z 519.1 [M + H]+. 160 ##STR00368## HCl salt, yellow solid;
.sup.1H-NMR (MeOD, 400 MHz): .delta. 8.83-8.87 (m, 2H), 7.94 (d, J
= 9.6 Hz, 1H), 7.85 (s, 1H), 7.84 (s, 1H), 7.59 (d, J = 6.8 Hz,
1H), 7.53 (s, 1H), 4.32 (t, J = 4.8 Hz, 2H), 4.17 (s, 3H), 4.11 (s,
3H), 3.81-3.86 (m, 1H), 3.49 (t, J = 4.8 Hz, 1H), 2.86 (s, 3H),
1.25-1.29 (m, 2H), 1.10-1.15 (m, 2H); LCMS: 96.4%, MS (ESI): m/z
506.2 [M + H]+. 161 ##STR00369## white solid; .sup.1H-NMR (MeOD,
400 MHz): .delta. 8.66 (d, J = 9.6 Hz, 1H), 8.48 (d, J = 5.2 Hz,
1H), 7.66 (s, 1H), 7.61 (d, J = 9.6 Hz, 1H), 7.37 (s, 1H), 7.31 (s,
1H), 6.95 (d, J = 5.6 Hz, 1H), 4.51 (t, J = 6.0 Hz, 3H), 4.22 (d, J
= 6.4 Hz, 2H), 3.92 (s, 3H), 3.85 (s, 3H), 3.56-3.66 (m, 1H),
3.35-3.53 (m, 1H), 1.17-1.30 (m, 1H), 1.01-1.12 (m, 2H), 0.96-0.99
(m, 2H); LCMS: 98.5%, MS (ESI): m/z 519.2 [M + H]+. 162
##STR00370## yellow solid; .sup.1H-NMR (MeOD, 400 MHz): .delta.
8.77 (d, J = 9.6 Hz, 1H), 8.58 (d, J = 5.2 Hz, 1H), 7.78 (s, 1H),
7.72 (d, J = 9.6 Hz, 1H), 7.49 (s, 1H), 7.42 (s, 1H), 7.07 (d, J =
5.2 Hz, 1H), 6.10- 6.42 (m, 1H), 4.29-4.34 (m, 2H), 4.04 (s, 3H),
3.97 (s, 3H), 3.77-3.97 (m, 1H), 1.36 (d, J = 6.4 Hz, 1H),
1.22-1.24 (m, 2H), 1.08-1.10 (m, 2H); LCMS: 99.1%, MS (ESI): m/z
513.2 [(M + H)]+. 163 ##STR00371## white solid; .sup.1H-NMR (MeOD,
400 MHz): .delta. 8.70 (d, J = 6.8 Hz, 1H), 8.54 (d, J = 8.8 Hz,
1H), 8.45 (d, J = 2.4 Hz, 1H), 7.93 (dd, J = 2.8, 9.2 Hz, 1H), 7.87
(s, 1H), 7.64 (s, 1H), 7.47 (s, 1H), 7.04 (d, J = 6.8 Hz, 1H),
4.06-4.19 (m, 11H), 2.87-2.94 (m, 1H), 2.19-2.22 (m, 2H), 1.99-2.05
(m, 4H), 1.21 (d, J = 6.0 Hz, 3H); HPLC: 96.0%, MS (ESI): m/z 534.3
[(M + H)]+. 164 ##STR00372## yellow solid; .sup.1H-NMR (MeOD, 400
MHz): .delta. 8.72 (d, J = 6.8 Hz, 1H), 8.56 (d, J = 9.2 Hz, 1H),
8.46 (d, J = 2.8 Hz, 1H), 7.96 (d, J = 2.8 Hz, 1H), 7.89 (s, 1H),
7.71 (s, 1H), 7.49 (s, 1H), 7.06 (d, J = 6.8 Hz, 1H), 4.14-4.18 (m,
5H), 4.12 (s, BH), 4.00-4.14 (m, 4H), 3.49-3.55 (m, 2H), 2.21-2.24
(m, 1H), 1.92-2.00 (m, 2H), 1.86-1.90 (m, 2H), 1.53-1.60 (m, 2H),
0.97 (t, J = 7.6 Hz, 3H); LCMS: 96.2%, MS (ESI): m/z 548.3 [(M +
H)]+. 165 ##STR00373## white powder; .sup.1H-NMR (DMSO-d6, 400
MHz): .delta. 10.29 (1H, brs), 8.83 (1H, d, J = 6.4 Hz), 8.13 (1H,
dd, J = 13.2, 2.0 Hz), 7.91 (1H, s), 7.85 (1H, d, J = 8.8 Hz), 7.76
(1H, s), 7.70 (1H, s), 7.54-7.61 (1H, m), 6.95 (1H, d, J = 6.0 Hz),
4.31-4.37 (2H, m), 4.12 (2H, t, J = 7.2 Hz), 4.05 (6H, s),
3.48-3.66 (2H, m), 2.90 (6H, d, J = 4.8 Hz), 1.80-1.92 (2H, m),
0.89 (3H, t, J = 7.2 Hz); LCMS: 95.7%, MS (ESI): m/z 538.2 [M +
H]+. 166 ##STR00374## white powder; .sup.1H-NMR (CDCl3, 400 MHz):
.delta. 9.84 (1H, brs), 8.72 (1H, d, J = 8.4 Hz), 8.51-8.57 (1H,
m), 8.31 (1H, s), 8.20 (1H, s), 7.63-7.72 (2H, m), 7.14 (1H, s),
6.72-6.79 (1H, m), 4.39 (2H, t, J = 7.2 Hz), 4.19 (3H, s), 4.13
(3H, s), 4.03 (2H, d, J = 6.4 Hz), 2.78-2.94 (3H, m), 2.14-2.23
(2H, m), 1.95-2.03 (4H, m); LCMS: 100%, MS (ESI): m/z 572.2 [M +
H]+. 167 ##STR00375## white solid; .sup.1H-NMR (CDCl3, 400 MHz):
.delta. 9.57 (1H, brs), 8.63 (1H, d, J = 5.2 Hz), 8.57 (1H, d, J =
9.2 Hz), 8.24 (1H, d, J = 2.8 Hz), 8.18 (1H, s), 7.51-7.59 (2H, m),
7.15 (1H, s), 6.46 (1H, d, J = 5.2 Hz), 4.06-4.11 (2H, m), 4.05
(3H, s), 4.01 (1H, d, J = 6.4 Hz), 2.85-2.95 (1H, m), 2.15-2.24
(2H, m), 1.88-2.02 (6H, m), 0.95 (3H, t, J = 7.2 Hz); LCMS: 100%,
MS (ESI): 572.1 m/z [(M + H)]+. 168 ##STR00376## HCl salt, yellow
solid; .sup.1H-NMR (MeOD, 400 MHz): .delta. 8.71 (d, J = 6.4 Hz,
1H), 8.53 (d, J = 9.2 Hz, 1H), 8.48 (d, J = 2.4 Hz, 1H), 7.97 (dd,
J = 2.4, 9.2 Hz, 1H), 7.90 (s, 1H), 7.86 (s, 1H), 7.50 (s, 1H),
7.04 (d, J = 6.8 Hz, 1H), 4.69 (q, J = 8.4 Hz, 2H), 4.61 (t, J =
5.2 Hz, 2H), 4.13 (s, 3H), 4.10 (s, 3H), 3.76 (t, J = 5.2 Hz, 2H),
2.87-2.89 (m, 2H), 0.96-0.99 (m, 4H); HPLC: 96.8%, MS (ESI): 573.2
m/z [(M + H)]+. 169 ##STR00377## white solid; .sup.1H-NMR (MeOD,
400 MHz): .delta. 8.43-8.46 (m, 2H), 8.32 (d, J = 2.8 Hz, 1H),
7.76-7.80 (m, 2H), 7.64 (s, 1H), 7.36 (s, 1H), 6.59 (d, J = 5.2 Hz,
1H), 4.64 (q, J = 8.4 Hz, 2H), 4.16 (d. J = 8.4 Hz, 2H), 4.01 (d. J
= 3.2 Hz, 6H), 2.66-2.85 (m, 1H), 2.62-2.65 (m, 3H), 2.42-2.45 (m,
1H), 2.38 (s, 1H), 2.02-2.04 (m, 1H), 1.63-1.66 (m, 1H); HPLC:
98.3%, MS (ESI): 544.2 m/z [(M + H)]+. 170 ##STR00378## yellow
solid; .sup.1H-NMR (MeOD, 400 MHz): .delta. 8.72 (d, J = 6.8 Hz,
1H), 8.50 (br s, 1H), 8.45 (d, J = 9.2 Hz, 1H), 8.06 (dd, J = 9.2,
2.8 Hz, 1H), 7.85 (s, 1H), 7.78 (s, 1H), 7.50 (s, 1H), 7.08 (d, J =
6.8 Hz, 1H), 4.31 (t, J = 6.0 Hz, 2H), 4.13 (s, 3H), 4.09 (s, 3H),
3.76-3.80 (m, 1H), 2.77-2.83 (m, 2H), 1.21-1.25 (m, 2H), 1.08- 1.11
(m, 2H); HPLC: 97.9%, MS (ESI): 544.2 m/z [(M + H)]+. 171
##STR00379## yellow solid; .sup.1H-NMR (MeOD, 400 MHz): .delta.
8.90 (d, J = 9.6 Hz, 1H), 8.79 (d, J = 6.8 Hz, 1H), 7.87 (d, J =
9.6 Hz, 1H), 7.84 (s, 1H), 7.76 (s, 1H), 7.52 (d, J = 6.8 Hz, 1H),
7.49 (s, 1H), 4.24- 4.26 (m, 2H), 4.14 (s, 3H), 4.09 (s, 3H),
3.79-3.81 (m, 3H), 3.43 (s, 3H), 1.30 (d, J = 6.4 Hz, 1H),
1.19-1.21 (m, 2H), 1.04- 1.10 (m, 2H); HPLC: 97.7%, MS (ESI): 507.2
m/z [(M + H)]+. 172 ##STR00380## HCl salt, yellow solid;
.sup.1H-NMR (MeOD, 400 MHz): .delta. 8.81-8.07 (m, 2H), 8.06 (d, J
= 9.6 Hz, 1H), 7.82 (d, J = 7.2 Hz, 2H), 7.66 (d, J = 6.8 Hz, 1H),
7.57 (s, 1H), 4.24-4.30 (m, 2H), 4.19-4.23 (m, 4H), 4.15 (s, 3H),
4.09 (s, 3H), 3.80-3.85 (m, 1H), 3.37- 3.38 (m, 1H), 1.24-1.27 (m,
2H), 1.07-1.12 (m, 2H); HPLC: 94.3%, MS (ESI): m/z 518.3 [M + H]+.
173 ##STR00381## yellow solid; .sup.1H-NMR (MeOD, 400 MHz): .delta.
8.89 (d, J = 10 Hz, 1H), 8.79 (d, J = 6.8 Hz, 1H), 7.87 (d, J = 9.6
Hz, 1H) 7.82 (s, 1H), 7.79 (s, 1H), 7.53 (d, J = 6.8 Hz, 1H), 7.48
(s, 1H), 4.33 (t, J = 5.6 Hz, 2H), 4.14 (s, 3H), 4.09 (s, 3H),
3.76-3.78 (m, 1H), 2.78-2.83 (m, 2H), 1.21-1.25 (m, 2H), 1.07-1.09
(m, 1H); HPLC: 98.6%, MS (ESI): m/z 545.2 [M + H]+. 174
##STR00382## white solid; .sup.1H-NMR (CDCl3, 400 MHz): .delta.
9.47 (s, 1H), 8.54 (d, J = 9.2 Hz), 8.51 (d, J = 5.2 Hz), 8.24 (d,
J = 2.8 Hz, 1H), 7.58-7.62 (m, 2H), 7.45 (s, 1H), 7.30 (s, 1H),
6.46 (d, J = 5.2 Hz, 1H), 4.32 (s, 1H), 4.20-4.23 (m, 3H), 4.09 (d,
6H), 3.43 (t, J = 7.6 Hz, 2H), 3.18 (t, J = 6.0 Hz, 2H), 2.96-3.03
(m, 1H), 2.50- 2.53 (m, 1H), 2.36 (s, 3H), 1.29 (d, J = 6.4 Hz,
3H); HPLC: 94.8%, MS (ESI): m/z 549.3 [M + H]+. 175 ##STR00383##
white solid; .sup.1H-NMR (MeOD, 400 MHz) .delta. 8.44-8.47 (m, 2H),
8.33 (d, J = 2.4 Hz, 1H), 7.78 (dd, J =8.8, 2.8 Hz, 1H), 7.70 (s,
1H), 7.65 (s, 1H), 7.38 (s, 1H), 6.60 (d, J = 5.6 Hz, 1H),
6.12- 6.42 (m, 1H), 4.30-4.38 (m, 2H), 4.15-4.22 (m, 2H), 4.05-4.09
(m, 1H), 4.02 (d, J = 3.6 Hz, 6H), 1.21 (d, J = 6.0 Hz, 3H); HPLC:
96.3%, MS (ESI): m/z 530.2 [M + H]+. 176 ##STR00384## white solid;
.sup.1H-NMR (MeOD, 400 MHz) .delta. 8.46 (1H, d, J = 5.2 Hz), 8.42
(1H, d, J = 9.2 Hz), 8.33 (1H, d, J = 2.8 Hz), 7.79 (1H, dd, J =
9.2, 2.8 Hz), 7.65 (1H, s), 7.38 (1H, s), 6.61 (1H, d, J = 5.2 Hz),
4.02 (6H, d, J = 4.0 Hz), 3.11 (2H, t, J = 7.6 Hz), 3.04 (2H, t, J
= 7.6 Hz), 2.72-2.83 (2H, m), 2.32-2.39 (1H, m), 2.03- 2.12 (3H,
m), 1.78-1.89 (4H, m), 1.63-1.72 (2H, m); LCMS: 99.1%, MS (ESI):
m/z 571.1 [M + H]+. 177 ##STR00385## HCl salt, yellow powder;
.sup.1H-NMR (DMSO-d6, 400 MHz): .delta. 9.63 (1H, brs), 8.86 (1H,
d, J = 6.8 Hz), 8.52 (1H, d, J = 2.8 Hz), 8.38 (1H, d, J = 8.8 Hz),
8.02 (1H, dd, J = 9.2, 2.8 Hz), 7.96 (1H, s), 7.79 (1H, s), 7.74
(1H, s), 7.02 (1H, d, J = 6.8 Hz), 4.38-4.43 (2H, m), 4.12 (2H, t,
J = 6.8 Hz), 4.02-4.07 (7H, m), 3.48-3.51 (1H, m, overlap with
water signal), 2.91 (6H, d, J = 4.8 Hz), 1.80-1.92 (2H, m), 0.89
(3H, t, J = 7.2 Hz); LCMS: 94.7%, MS (ESI): m/z 521.2 [M + H]+. 178
##STR00386## white powder; .sup.1H-NMR (400 MHz, CDCl3) .delta.
12.37 (1H, s), 8.51 (1H, d, J = 5.2 Hz), 8.44-8.48 (2H, m), 8.31
(1H, d, J = 2.8 Hz), 7.54-7.61 (2H, m), 7.44 (1H, s), 7.37 (1H, s),
6.46 (1H, d, J = 5.2 Hz), 4.74 (2H, q, J = 8.4 Hz), 4.06 (6H, d, J
= 1.6 Hz), 1.74-1.88 (1H, m), 0.93-1.09 (2H, m), 0.57-0.75 (2H, m);
LCMS: 99.7%, MS (ESI): m/z 541.1 [M + H]+. 179 ##STR00387## white
powder; .sup.1H-NMR (CDCl3, 400 MHz): .delta. 9.37 (1H, s), 8.51
(1H, d, J = 5.6 Hz), 8.41 (1H, d, J = 8.8 Hz), 8.25 (1H, d, J = 2.8
Hz), 7.52-7.58 (2H, m), 7.43 (1H, s), 6.59 (1H, s), 6.46 (1H, d, J
= 5.2 Hz), 4.60 (2H, q, J = 8.0 Hz), 4.06 (3H, s), 4.06 (3H, s),
2.05-2.15 (1H, m), 1.09-1.19 (2H, m), 0.79-0.86 (2H, m); LCMS:
100%, MS (ESI): m/z 546.0 [M + H]+ .
* * * * *